text:
Contact Us
Enter Your Search:
Search tips
Search criteria
Journal titles
Results 1-25 (1361244)
« Previous
Go to page number:
Next »
Clipboard (0)
Related Articles
A single nucleotide polymorphism in the p27Kip1 gene is associated with primary patency of lower extremity vein bypass grafts
Conte, Michael S.
Owens, Christopher D.
Belkin, Michael
Creager, Mark A.
Edwards, Karen L.
Gasper, Warren J.
Kenagy, Richard D.
LeBoeuf, Renee C.
Sobel, Michael
Clowes, Alexander
Journal of vascular surgery
Factors responsible for the variability in outcomes after lower extremity vein bypass grafting (LEVBG) are poorly understood. Recent evidence has suggested that a single nucleotide polymorphism (SNP) in the promoter region of the p27Kip1 gene, a cell-cycle regulator, is associated with coronary in-stent restenosis. We hypothesized an association with vein graft patency.
This was a retrospective genetic association study nested within a prospective cohort of 204 patients from three referral centers undergoing LEVBG for claudication or critical ischemia. The main outcome measure was primary vein graft patency.
All patients were followed up for a minimum of 1 year with duplex graft surveillance (median follow-up, 893 days; interquartile range, 539-1315). Genomic DNA was isolated and SNP analysis for the p27Kip1-838C>A variants was performed. Allele frequencies were correlated with graft outcome using survival analysis and Cox proportional hazards modeling. The p27Kip1-838C>A allele frequencies observed were CA, 53%; CC, 30%; and AA, 17%, satisfying Hardy-Weinberg equilibrium. Race (P = .025) and history of coronary artery disease (P = .027) were different across the genotypes; all other baseline variables were similar. Primary graft patency was greater among patients with the −838AA genotype (75% AA vs 55% CA/CC at 3 years; P = .029). In a Cox proportional hazards model including age, sex, race, diabetes, critical limb ischemia, redo (vs primary) bypass, vein type, and baseline C-reactive protein level, the p27Kip1-838AA genotype was significantly associated with higher graft patency (hazard ratio for failure, 0.4; 95% confidence interval, 0.17-0.93). Genotype was also associated with early (0-1 month) changes in graft lumen diameter by ultrasound imaging.
These data suggest that the p27Kip1-838C>A SNP is associated with LEVBG patency and, together with previous reports, underscore a central role for p27Kip1 in the generic response to vascular injury.
Related Articles
A new tumour suppression mechanism by p27Kip1: EGFR down-regulation mediated by JNK/c-Jun pathway inhibition
Fang, Yong
Wang, Yihong
Wang, Yulei
Meng, Yan
Zhu, Junlan
Jin, Honglei
Li, Jingxia
Zhang, Dongyun
Yu, Yonghui
Wu, Xue-Ru
Huang, Chuanshu
Biochemical Journal
2014;463(Pt 3):383-392.
p27Kip1 is a potent inhibitor of cyclin-dependent kinases that drive G1-to-S cell-cycle transition. Reduced p27Kip1 expression is prevalent in a wide range of human tumours; however, the exact mechanism(s) of p27Kip1-mediated tumour suppression remains obscure. In the present study, we identified a close inverse relationship between p27Kip1 and EGFR (epidermal growth factor receptor) expression: the parental T24 human bladder cancer cells had high p27Kip1 expression but low EGFR expression and, in striking contrast, the metastatic derivative of T24 (T24T) had low p27Kip1 expression but high EGFR expression. This relationship was also found in various human cancer tissues, and was not only just correlative but also causal; depletion of p27Kip1 in MEF (mouse embryonic fibroblast) cells resulted in markedly elevated EGFR expression, a result reproducible with an Egfr promoter-luciferase reporter in both T24 and MEF cells, suggesting transcriptional repression of EGFR by p27Kip1. Indeed, p27Kip1 was found to regulate EGFR expression via the JNK (c-Jun N-terminal kinase)/c-Jun transcription factor: p27Kip1 deficiency activated JNK/c-Jun, whereas inhibition of JNK/c-Jun by dominant-negative mutants dramatically repressed Egfr transcription. Furthermore, the proximal promoter of the Egfr gene was crucial for its transcription, where the recruiting activity of c-Jun was much greater in p27Kip1−/− cells than in p27Kip1+/+ cells. Introduction of GFP–p27Kip1 into T24T cells suppressed JNK/c-Jun activation, EGFR expression and anchorage-independent growth. The results of the present study demonstrate that p27Kip1 suppresses JNK/c-Jun activation and EGFR expression in MEFs and human bladder cancer cells, and the results obtained are consistent with those from human cancer specimens. The present study provides new insights into p27Kip1 suppression of cancer cell growth, migration and metastasis.
An inverse relationship between p27Kip1 and EGFR expression in parental T24 human bladder cancer cells and various human cancer tissues was found. Depletion of p27Kip1 in cells markedly elevated EGFR expression through transcriptional repression of Egfr by p27Kip1 via the JNK/c-Jun cascade.
bladder cancer; c-Jun N-terminal kinase (JNK)/c-Jun pathway; epidermal growth factor receptor (EGFR); p27Kip1; signal transduction pathway; AP-1, activator protein 1; BME, basal medium Eagle; CDK, cyclin-dependent kinase; DMEM, Dulbecco's modified Eagle's medium; EGFR, epidermal growth factor receptor; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HSF-1, heat-shock factor 1; Hsp, heat-shock protein; IHC, immunohistochemistry; JNK, c-Jun N-terminal kinase; MEF, mouse embryonic fibroblast; RT, reverse transcription; SP1, specificity protein 1
Related Articles
The major transcription initiation site of the p27Kip1 gene is conserved in human and mouse and produces a long 5'-UTR
Coleman, Jennifer
Hawkinson, Michelle
Miskimins, Robin
Miskimins, W Keith
BMC Molecular Biology
The cyclin-dependent kinase inhibitor p27Kip1 is essential for proper control of cell cycle progression. The levels of p27Kip1 are regulated by several mechanisms including transcriptional and translational controls. In order to delineate the molecular details of these regulatory mechanisms it is important to identify the transcription initiation site within the p27Kip1 gene, thereby defining the promoter region of the gene and the 5'-untranslated region of the p27Kip1 mRNA. Although several previous studies have attempted to map p27Kip1 transcription start sites, the results vary widely for both the mouse and human genes. In addition, even though the mouse and human p27Kip1 gene sequences are very highly conserved, the reported start sites are notably different.
In this report, using a method that identifies capped ends of mRNA molecules together with RNase protection assays, we demonstrate that p27Kip1 transcription is initiated predominantly from a single site which is conserved in the human and mouse genes. Initiation at this site produces a 5'-untranslated region of 472 nucleotides in the human p27Kip1 mRNA and 502 nucleotides in the mouse p27Kip1 mRNA. In addition, several minor transcription start sites were identified for both the mouse and human genes.
These results demonstrate that the major transcription initiation sites in the mouse and human p27Kip1 genes are conserved and that the 5'-UTR of the p27Kip1 mRNA is much longer than generally believed. It will be important to consider these findings when designing experiments to identify elements that are involved in regulating the cellular levels of p27Kip1.
Related Articles
Effect of hypoxia and Beraprost sodium on human pulmonary arterial smooth muscle cell proliferation: the role of p27kip1
Kadowaki, Maiko
Mizuno, Shiro
Demura, Yoshiki
Ameshima, Shingo
Miyamori, Isamu
Ishizaki, Takeshi
Respiratory Research
Hypoxia induces the proliferation of pulmonary arterial smooth muscle cell (PASMC) in vivo and in vitro, and prostacyclin analogues are thought to inhibit the growth of PASMC. Previous studies suggest that p27kip1, a kind of cyclin-dependent kinase inhibitor, play an important role in the smooth muscle cell proliferation. However, the mechanism of hypoxia and the subcellular interactions between p27kip1 and prostacyclin analogues in human pulmonary arterial smooth muscle cell (HPASMC) are not fully understood.
We investigated the role of p27kip1 in the ability of Beraprost sodium (BPS; a stable prostacyclin analogue) to inhibit the proliferation of HPASMC during hypoxia. To clarify the biological effects of hypoxic air exposure and BPS on HPASMC, the cells were cultured in a hypoxic chamber under various oxygen concentrations (0.1–21%). Thereafter, DNA synthesis was measured as bromodeoxyuridine (BrdU) incorporation, the cell cycle was analyzed by flow cytometry with propidium iodide staining. The p27kip1 mRNA and protein expression and it's stability was measured by real-time RT-PCR and Western blotting. Further, we assessed the role of p27kip1 in HPASMC proliferation using p27kip1 gene knockdown using small interfering RNA (siRNA) transfection.
Although severe hypoxia (0.1% oxygen) suppressed the proliferation of serum-stimulated HPASMC, moderate hypoxia (2% oxygen) enhanced proliferation in accordance with enhanced p27kip1 protein degradation, whereas BPS suppressed HPASMC proliferation under both hypoxic and normoxic conditions by suppressing p27kip1 degradation with intracellular cAMP-elevation. The 8-bromo-cyclic adenosine monophosphate (8-Br-cAMP), a cAMP analogue, had similar action as BPS in the regulation of p27kip1. Moderate hypoxia did not affect the stability of p27kip1 protein expression, but PDGF, known as major hypoxia-induced growth factors, significantly decreased p27kip1 protein stability. We also demonstrated that BPS and 8-Br-cAMP suppressed HPASMC proliferation under both hypoxic and normoxic conditions by blocking p27kip1 mRNA degradation. Furthermore, p27kip1 gene silencing partially attenuated the effects of BPS and partially restored hypoxia-induced proliferation.
Our study suggests that moderate hypoxia induces HPASMC proliferation, which is partially dependent of p27kip1 down-regulation probably via the induction of growth factors such as PDGF, and BPS inhibits both the cell proliferation and p27kip1 mRNA degradation through cAMP pathway.
Related Articles
Streptogramin- and tetracycline-responsive dual regulated expression of p27Kip1 sense and antisense enables positive and negative growth control of Chinese hamster ovary cells
Fux, Cornelia
Moser, Samuel
Schlatter, Stefan
Rimann, Markus
Bailey, James E.
Fussenegger, Martin
Nucleic Acids Research
We constructed a dual regulated expression vector cassette (pDuoRex) whereby two heterologous genes can be independently regulated via streptogramin- and tetracycline-responsive promoters. Two different constructs containing growth-promoting and growth-inhibiting genes were stably transfected in recombinant Chinese hamster ovary (CHO) cells that express the streptogramin- and tetracycline-dependent transactivators in a dicistronic configuration. An optimally balanced heterologous growth control scenario was achieved by reciprocal expression of the growth-inhibiting human cyclin-dependent kinase inhibitor p27Kip1 in sense (p27Kip1S) and antisense (p27Kip1AS) orientation. Exclusive expression of p27Kip1S resulted in complete G1-phase-specific growth arrest, while expression of only p27Kip1AS showed significantly increased proliferation compared to control cultures (both antibiotics present), presumably by decreasing host cell p27Kip1 expression. In a second system, a derivative of pDuoRex encoding streptogramin-responsive expression of the growth-promoting SV40 small T antigen (sT) and tetracycline-regulated expression of p27Kip1 was stably transfected into CHO cells. Expression of sT alone resulted in an increase in cell proliferation, but the expression of p27Kip1 failed to provide the expected G1-specific growth arrest despite having demonstrated expression of the protein. This illustrates the difficulty in balancing the complex pathways underlying cell proliferation control through the expression of two functionally distinct genes involved in those pathways, and how a single-gene sense/antisense approach using pDuoRex can overcome this barrier to complete metabolic engineering control.
Related Articles
Cortactin Modulates RhoA Activation and Expression of Cip/Kip Cyclin-Dependent Kinase Inhibitors To Promote Cell Cycle Progression in 11q13-Amplified Head and Neck Squamous Cell Carcinoma Cells ▿ †
Croucher, David R.
Rickwood, Danny
Tactacan, Carole M.
Musgrove, Elizabeth A.
Daly, Roger J.
Molecular and Cellular Biology
The cortactin oncoprotein is frequently overexpressed in head and neck squamous cell carcinoma (HNSCC), often due to amplification of the encoding gene (CTTN). While cortactin overexpression enhances invasive potential, recent research indicates that it also promotes cell proliferation, but how cortactin regulates the cell cycle machinery is unclear. In this article we report that stable short hairpin RNA-mediated cortactin knockdown in the 11q13-amplified cell line FaDu led to increased expression of the Cip/Kip cyclin-dependent kinase inhibitors (CDKIs) p21WAF1/Cip1, p27Kip1, and p57Kip2 and inhibition of S-phase entry. These effects were associated with increased binding of p21WAF1/Cip1 and p27Kip1 to cyclin D1- and E1-containing complexes and decreased retinoblastoma protein phosphorylation. Cortactin regulated expression of p21WAF1/Cip1 and p27Kip1 at the transcriptional and posttranscriptional levels, respectively. The direct roles of p21WAF1/Cip1, p27Kip1, and p57Kip2 downstream of cortactin were confirmed by the transient knockdown of each CDKI by specific small interfering RNAs, which led to partial rescue of cell cycle progression. Interestingly, FaDu cells with reduced cortactin levels also exhibited a significant diminution in RhoA expression and activity, together with decreased expression of Skp2, a critical component of the SCF ubiquitin ligase that targets p27Kip1 and p57Kip2 for degradation. Transient knockdown of RhoA in FaDu cells decreased expression of Skp2, enhanced the level of Cip/Kip CDKIs, and attenuated S-phase entry. These findings identify a novel mechanism for regulation of proliferation in 11q13-amplified HNSCC cells, in which overexpressed cortactin acts via RhoA to decrease expression of Cip/Kip CDKIs, and highlight Skp2 as a downstream effector for RhoA in this process.
Related Articles
p27KIP1 Deletions in Childhood Acute Lymphoblastic Leukemia
Komuro, Hiroaki
Valentine, Marcus B
Rubnitz, Jeffrey E
Saito, Midori
Raimondi, Susana C
Carroll, Andrew J
Yi, Taolin
Sherr, Charles J
Look, A Thomas
Neoplasia (New York, N.Y.)
The p27KIP1 gene, which encodes a cyclin-dependent kinase (CDK) inhibitor, has been assigned to chromosome band 12p12, a region often affected by cytogenetically apparent deletions or translocations in childhood acute lymphoblastic leukemia (ALL). As described here, fluorescence in situ hybridization (FISH) analysis of 35 primary ALL samples with cytogenetic evidence of 12p abnormalities revealed hemizygous deletions of p27KIP1 in 29 cases. Further analysis of 19 of these cases with two additional gene-specific probes from the 12p region (hematopoietic cell phosphatase, HCP and cyclin D2, CCND2) showed that p27KIP1 is located more proximally on the short arm of chromosome 12 and is deleted more frequently than either HCP or CCND2. Of 16 of these cases with hemizygous deletion of p27KIP1, only eight showed loss of HCP or CCND2, whereas loss of either of the latter two loci was uniformly associated with loss of p27KIP1. Missense mutations or mutations leading to premature termination codons were not detected in the coding sequences of the retained p27KIP1 alleles in any of the 16 ALL cases examined, indicating a lack of homozygous inactivation. By Southern blot analysis, one case of primary T-cell ALL had hemizygous loss of a single p27KIP1 allele and a 34.5-kb deletion, including the second coding exon of the other allele. Despite homozygous inactivation of p27KIP1 in this case, our data suggest that haploinsufficiency for p27KIP1 is the primary consequence of 12p chromosomal deletions in childhood ALL. The oncogenic role of reduced, but not absent, levels of p27KIP1 is supported by recent studies in murine models and evidence that this protein not only inhibits the activity of complexes containing CDK2 and cyclin E, but also promotes the assembly and catalytic activity of CDK4 or CDK6 in complexes with cyclin D.
acute lymphoblastic leukemia; p27KIP1; cell cycle inhibitor
Related Articles
A Novel Mutation in the Upstream Open Reading Frame of the CDKN1B Gene Causes a MEN4 Phenotype
Occhi, Gianluca
Regazzo, Daniela
Trivellin, Giampaolo
Boaretto, Francesca
Ciato, Denis
Bobisse, Sara
Ferasin, Sergio
Cetani, Filomena
Pardi, Elena
Korbonits, Márta
Pellegata, Natalia S.
Sidarovich, Viktoryia
Quattrone, Alessandro
Opocher, Giuseppe
Mantero, Franco
Scaroni, Carla
PLoS Genetics
The CDKN1B gene encodes the cyclin-dependent kinase inhibitor p27KIP1, an atypical tumor suppressor playing a key role in cell cycle regulation, cell proliferation, and differentiation. Impaired p27KIP1 expression and/or localization are often observed in tumor cells, further confirming its central role in regulating the cell cycle. Recently, germline mutations in CDKN1B have been associated with the inherited multiple endocrine neoplasia syndrome type 4, an autosomal dominant syndrome characterized by varying combinations of tumors affecting at least two endocrine organs. In this study we identified a 4-bp deletion in a highly conserved regulatory upstream ORF (uORF) in the 5′UTR of the CDKN1B gene in a patient with a pituitary adenoma and a well-differentiated pancreatic neoplasm. This deletion causes the shift of the uORF termination codon with the consequent lengthening of the uORF–encoded peptide and the drastic shortening of the intercistronic space. Our data on the immunohistochemical analysis of the patient's pancreatic lesion, functional studies based on dual-luciferase assays, site-directed mutagenesis, and on polysome profiling show a negative influence of this deletion on the translation reinitiation at the CDKN1B starting site, with a consequent reduction in p27KIP1 expression. Our findings demonstrate that, in addition to the previously described mechanisms leading to reduced p27KIP1 activity, such as degradation via the ubiquitin/proteasome pathway or non-covalent sequestration, p27KIP1 activity can also be modulated by an uORF and mutations affecting uORF could change p27KIP1 expression. This study adds the CDKN1B gene to the short list of genes for which mutations that either create, delete, or severely modify their regulatory uORFs have been associated with human diseases.
Author Summary
Gene expression can be modulated at different steps on the way from DNA to protein including control of transcription, translation, and post-translational modifications. An abnormality in the regulation of mRNA and protein expression is a hallmark of many human diseases, including cancer. In some eukaryotic genes translation can be influenced by small DNA sequences termed upstream open reading frames (uORFs). These elements located upstream to the gene start codon may either negatively influence the ability of the translational machinery to reinitiate translation of the main protein or, much less frequently, stimulate protein translation by enabling the ribosomes to bypass cis-acting inhibitory elements. CDKN1B, which encodes the cell cycle inhibitor p27KIP1, includes an uORF in its 5′UTR sequence. p27KIP1 expression is often reduced in cancer, and germline mutations have been identified in CDKN1B in patients affected with a syndrome (MEN4) characterized by varying combinations of tumors in endocrine glands. Here we show that a small deletion in the uORF upstream to CDKN1B reduces translation reinitiation efficiency, leading to underexpression of p27KIP1 and coinciding with tumorigenesis. This study describes a novel mechanism by which p27KIP1 could be underexpressed in human tumors. In addition, our data provide a new insight to the unique pathogenic potential of uORFs in human diseases.
Related Articles
S-phase kinase-associated protein-2 (Skp2) promotes vascular smooth muscle cell proliferation and neointima formation in vivo
Wu, Yih-Jer
Sala-Newby, Graciela B.
Shu, Kuo-Tung
Yeh, Hung-I.
Nakayama, Keiichi I.
Nakayama, Keiko
Newby, Andrew C.
Bond, Mark
Journal of Vascular Surgery
Vascular smooth muscle cell (VSMC) proliferation plays an important role in the development of postangioplasty or in-stent restenosis, venous graft failure, and atherosclerosis. Our previous work has demonstrated S-phase kinase-associated protein-2 (Skp2), an F-box subunit of SCFSkp2 ubiquitin ligase, as an important mediator and common final pathway for growth factors, extracellular matrices, and cyclic-nucleotides to regulate VSMC proliferation in vitro. However, whether alteration of Skp2 function also regulates VSMC proliferation in vivo and neointimal thickening postvascular injury remains unclear. We investigated the effect of Skp2 on VSMC proliferation and neointimal formation in vivo.
Methods and Results
Firstly, we demonstrated that Skp2-null mice developed significantly smaller neointimal areas than wild-type mice after carotid ligation. Secondly, to further identify a local rather than a systemic effect of Skp2 alteration, we demonstrated that adenovirus-mediated expression of dominant-negative Skp2 in the balloon-injured rat carotid artery significantly increased medial p27Kip1 levels, inhibited VSMC proliferation, and the subsequent neointimal thickening. Lastly, to determine if Skp2 alone is sufficient to drive VSMC proliferation and lesion development in vivo, we demonstrated that adenovirus-delivery of wild-type Skp2 to the minimally-injured rat carotids is sufficient to downregulate p27Kip1 protein levels, enhanced medial VSMC proliferation, and the neointimal thickening.
This data provides, we believe for the first time, a more comprehensive understanding of Skp2 in the regulation of VSMC proliferation and neointimal formation and suggests that Skp2 is a promising target in the treatment of vasculoproliferative diseases.
Clinical Relevance
This manuscript describes our latest work investigating the role of the Skp2, an F-box protein component of the SCFskp2 ubiquitin-ligase, in promoting VSMC proliferation, and neointima formation in response to vascular injury in vivo. Our previous work has identified a major role for Skp2 as a key target for numerous positive and negative growth regulatory signals in vitro. These signals converge to regulate the expression of Skp2, which then controls cell-cycle progression by promoting degradation of the cyclin-dependent kinase inhibitor, p27Kip1. Until now, there has been no data in the literature on the role played by Skp2 in the regulation of VSMC proliferation and neointima formation in vivo. Our current manuscript describes, we believe for the first time, the important role played by Skp2 in these processes, using both mouse and rat arterial injury models. This is important because proliferation of VSMCs underlies the development of postangioplasty or post-stenting restenosis, venous graft failure, and transplant arteriosclerosis. Our work demonstrates for the first time that Skp2 is a major regulator of VSMC proliferation and neointimal thickening in vivo in response to vascular injury and highlights Skp2 as a potential target for future strategies designed to combat vasculoproliferative diseases.
Related Articles
ErbB2 Potentiates Breast Tumor Proliferation through Modulation of p27Kip1-Cdk2 Complex Formation: Receptor Overexpression Does Not Determine Growth Dependency
Lane, Heidi A.
Beuvink, Iwan
Motoyama, Andrea B.
Daly, John M.
Neve, Richard M.
Hynes, Nancy E.
Molecular and Cellular Biology
Overexpression of the ErbB2 receptor, a major component of the ErbB receptor signaling network, contributes to the development of a number of human cancers. ErbB2 presents itself, therefore, as a target for antibody-mediated therapies. In this respect, anti-ErbB2 monoclonal antibody 4D5 specifically inhibits the growth of tumor cells overexpressing ErbB2. We have analyzed the effect of 4D5-mediated ErbB2 inhibition on the cell cycle of the breast tumor cell line BT474. 4D5 treatment of BT474 cells resulted in a G1 arrest, preceded by rapid dephosphorylation of ErbB2, inhibition of cytoplasmic signal transduction pathways, accumulation of the cyclin-dependent kinase inhibitor p27Kip1, and inactivation of cyclin-Cdk2 complexes. Time courses demonstrated that 4D5 treatment redirects p27Kip1 onto Cdk2 complexes, an event preceding increased p27Kip1 expression; this correlates with the downregulation of c-Myc and D-type cyclins (proteins involved in p27Kip1 sequestration) and the loss of p27Kip1 from Cdk4 complexes. Similar events were observed in ErbB2-overexpressing SKBR3 cells, which exhibited reduced proliferation in response to 4D5 treatment. Here, p27Kip1 redistribution resulted in partial Cdk2 inactivation, consistent with a G1 accumulation. Moreover, p27Kip1 protein levels remained constant. Antisense-mediated inhibition of p27Kip1 expression in 4D5-treated BT474 cells further demonstrated that in the absence of p27Kip1 accumulation, p27Kip1 redirection onto Cdk2 complexes is sufficient to inactivate Cdk2 and establish the G1 block. These data suggest that ErbB2 overexpression leads to potentiation of cyclin E-Cdk2 activity through regulation of p27Kip1 sequestration proteins, thus deregulating the G1/S transition. Moreover, through comparison with an ErbB2-overexpressing cell line insensitive to 4D5 treatment, we demonstrate the specificity of these cell cycle events and show that ErbB2 overexpression alone is insufficient to determine the cellular response to receptor inhibition.
Related Articles
The Cyclin-Dependent Kinase Inhibitor p57Kip2 Regulates Cell Cycle Exit, Differentiation, and Migration of Embryonic Cerebral Cortical Precursors
Tury, Anna
Mairet-Coello, Georges
DiCicco-Bloom, Emanuel
Cerebral Cortex (New York, NY)
Mounting evidence indicates cyclin-dependent kinase (CDK) inhibitors (CKIs) of the Cip/Kip family, including p57Kip2 and p27Kip1, control not only cell cycle exit but also corticogenesis. Nevertheless, distinct activities of p57Kip2 remain poorly defined. Using in vivo and culture approaches, we show p57Kip2 overexpression at E14.5–15.5 elicits precursor cell cycle exit, promotes transition from proliferation to neuronal differentiation, and enhances process outgrowth, while opposite effects occur in p57Kip2-deficient precursors. Studies at later ages indicate p57Kip2 overexpression also induces precocious glial differentiation, suggesting stage-dependent effects. In embryonic cortex, p57Kip2 overexpression advances cell radial migration and alters postnatal laminar positioning. While both CKIs induce differentiation, p57Kip2 was twice as effective as p27Kip1 in inducing neuronal differentiation and was not permissive to astrogliogenic effects of ciliary neurotrophic factor, suggesting that the CKIs differentially modulate cell fate decisions. At molecular levels, although highly conserved N-terminal regions of both CKIs elicit cycle withdrawal and differentiation, the C-terminal region of p57Kip2 alone inhibits in vivo migration. Furthermore, p57Kip2 effects on neurogenesis and gliogenesis require the N-terminal cyclin/CDK binding/inhibitory domains, while previous p27Kip1 studies report cell cycle-independent functions. These observations suggest p57Kip2 coordinates multiple stages of corticogenesis and exhibits distinct and common activities compared with related family member p27Kip1.
gliogenesis; in utero electroporation; neurite outgrowth; neurogenesis; transfection
Related Articles
Vitamin E δ-Tocotrienol Induces p27Kip1-Dependent Cell-Cycle Arrest in Pancreatic Cancer Cells via an E2F-1-Dependent Mechanism
Hodul, Pamela J.
Dong, Yanbin
Husain, Kazim
Pimiento, Jose M.
Chen, Jiandong
Zhang, Anying
Francois, Rony
Pledger, Warren J.
Coppola, Domenico
Sebti, Said M.
Chen, Dung-Tsa
Malafa, Mokenge P.
PLoS ONE
Vitamin E δ-tocotrienol has been shown to have antitumor activity, but the precise molecular mechanism by which it inhibits the proliferation of cancer cells remains unclear. Here, we demonstrated that δ-tocotrienol exerted significant cell growth inhibition pancreatic ductal cancer (PDCA) cells without affecting normal human pancreatic ductal epithelial cell growth. We also showed that δ-tocotrienol-induced growth inhibition occurred concomitantly with G1 cell-cycle arrest and increased p27Kip1 nuclear accumulation. This finding is significant considering that loss of nuclear p27Kip1 expression is a well-established adverse prognostic factor in PDCA. Furthermore, δ-tocotrienol inactivated RAF-MEK-ERK signaling, a pathway known to suppress p27Kip1 expression. To determine whether p27Kip1 induction is required for δ-tocotrienol inhibition of PDCA cell proliferation, we stably silenced the CDKN1B gene, encoding p27Kip1, in MIAPaCa-2 PDCA cells and demonstrated that p27Kip1 silencing suppressed cell-cycle arrest induced by δ-tocotrienol. Furthermore, δ-tocotrienol induced p27Kip1 mRNA expression but not its protein degradation. p27Kip1 gene promoter activity was induced by δ-tocotrienol through the promoter's E2F-1 binding site, and this activity was attenuated by E2F-1 depletion using E2F-1 small interfering RNA. Finally, decreased proliferation, mediated by Ki67 and p27Kip1 expression by δ-tocotrienol, was confirmed in vivo in a nude mouse xenograft pancreatic cancer model. Our findings reveal a new mechanism, dependent on p27Kip1 induction, by which δ-tocotrienol can inhibit proliferation in PDCA cells, providing a new rationale for p27Kip1 as a biomarker for δ-tocotrienol efficacy in pancreatic cancer prevention and therapy.
Related Articles
AMPKα2 Deletion Exacerbates Neointima Formation by Upregulating Skp2 in Vascular Smooth Muscle Cells
Song, Ping
Wang, Shuangxi
He, Chaoyong
Wang, Shaobin
Liang, Bin
Viollet, Benoit
Zou, Ming-Hui
Circulation research
Adenosine monophosphate-activated protein kinase (AMPK), a metabolic and redox sensor, is reported to suppress cell proliferation of non-malignant and tumor cells. Whether AMPKα alters vascular neointima formation induced by vascular injury is unknown.
The aim of this study was to determine the roles of AMPKα in the development of vascular neointima hyperplasia and to elucidate the underlying mechanisms.
Methods and Results
Vascular smooth muscle cells (VSMCs) proliferation and neointimal hyperplasia were evaluated in cultured VSMCs and wire-injured mouse carotid arteries from wild-type (WT, C57BL/6J), AMPKα2−/−, and AMPKα1−/− VSMCs. Mouse VSMCs derived from aortas of AMPKα2−/− mice exhibited increased proliferation compared to either WT or AMPKα1−/− VSMCs. Further, deletion of AMPKα2, but not AMPKα1, reduced the level of p27Kip1, acyclin-dependent kinase inhibitor, and increased the level of S-phase kinase-associated protein 2 (Skp2), a known E3 ubiquitin ligase for p27Kip1, via activation of p52 nuclear factor kappa B (NF-κB)-2. Moreover, either pharmacological (i.e., via compound C) or genetical (i.e., via AMPKα2-specific siRNA) inhibition of AMPK decreased p27Kip1 levels, but increased the abundance of Skp2 in human VSMCs. Furthermore, gene silencing of Skp2 reversed the levels of p27Kip1 and VSMCs proliferation. Finally, neointima formation after mechanical arterial injury was increased in AMPKα2−/−, but not AMPKα1−/−, mice.
These findings indicate that deletion of AMPKα2 via p52-Skp2-mediated ubiquintination and degradation of p27Kip1 accentuates neointimal hyperplasia in response to wire injury.
Neointima formation; AMPK; VSMC; NF-κB; Skp2
Related Articles
Phosphorylation and Subcellular Localization of p27Kip1 Regulated by Hydrogen Peroxide Modulation in Cancer Cells
Ibañez, Irene L.
Bracalente, Candelaria
Notcovich, Cintia
Tropper, Ivanna
Molinari, Beatriz L.
Policastro, Lucía L.
Durán, Hebe
PLoS ONE
The Cyclin-dependent kinase inhibitor 1B (p27Kip1) is a key protein in the decision between proliferation and cell cycle exit. Quiescent cells show nuclear p27Kip1, but this protein is exported to the cytoplasm in response to proliferating signals. We recently reported that catalase treatment increases the levels of p27Kip1 in vitro and in vivo in a murine model. In order to characterize and broaden these findings, we evaluated the regulation of p27Kip1 by hydrogen peroxide (H2O2) in human melanoma cells and melanocytes. We observed a high percentage of p27Kip1 positive nuclei in melanoma cells overexpressing or treated with exogenous catalase, while non-treated controls showed a cytoplasmic localization of p27Kip1. Then we studied the levels of p27Kip1 phosphorylated (p27p) at serine 10 (S10) and at threonine 198 (T198) because phosphorylation at these sites enables nuclear exportation of this protein, leading to accumulation and stabilization of p27pT198 in the cytoplasm. We demonstrated by western blot a decrease in p27pS10 and p27pT198 levels in response to H2O2 removal in melanoma cells, associated with nuclear p27Kip1. Melanocytes also exhibited nuclear p27Kip1 and lower levels of p27pS10 and p27pT198 than melanoma cells, which showed cytoplasmic p27Kip1. We also showed that the addition of H2O2 (0.1 µM) to melanoma cells arrested in G1 by serum starvation induces proliferation and increases the levels of p27pS10 and p27pT198 leading to cytoplasmic localization of p27Kip1. Nuclear localization and post-translational modifications of p27Kip1 were also demonstrated by catalase treatment of colorectal carcinoma and neuroblastoma cells, extending our findings to these other human cancer types. In conclusion, we showed in the present work that H2O2 scavenging prevents nuclear exportation of p27Kip1, allowing cell cycle arrest, suggesting that cancer cells take advantage of their intrinsic pro-oxidant state to favor cytoplasmic localization of p27Kip1.
Related Articles
p21cip1 Degradation in Differentiated Keratinocytes Is Abrogated by Costabilization with Cyclin E Induced by Human Papillomavirus E7
Noya, Francisco
Chien, Wei-Ming
Broker, Thomas R.
Chow, Louise T.
Journal of Virology
The human papillomavirus (HPV) E7 protein promotes S-phase reentry in a fraction of postmitotic, differentiated keratinocytes. Here we report that these cells contain an inherent mechanism that opposes E7-induced DNA replication. In organotypic raft cultures of primary human keratinocytes, neither cyclin E nor p21cip1 is detectable in situ. However, E7-transduced differentiated cells not in S phase accumulate abundant cyclin E and p21cip1. We show that normally p21cip1 protein is rapidly degraded by proteasomes. In the presence of E7 or E6/E7, p21cip1, cyclin E, and cyclin E2 proteins were all up-regulated. The accumulation of p21cip1 protein is a posttranscriptional event, and ectopic cyclin E expression was sufficient to trigger it. In constract, cdk2 and p27kip1 were abundant in normal differentiated cells and were not significantly affected by E7. Cyclin E, cdk2, and p21cip1 or p27kip1 formed complexes, and relatively little kinase activity was found associated with cyclin E or cdk2. In patient papillomas and E7 raft cultures, all p27kip1-positive cells were negative for bromodeoxyuridine (BrdU) incorporation, but only some also contained cyclin E and p21cip1. In contrast, all cyclin E-positive cells also contained p27kip1. When the expression of p21cip1 was reduced by rottlerin, a PKC δ inhibitor, p27kip1- and BrdU-positive cells remained unchanged. These observations show that high levels of endogenous p27kip1 can prevent E7-induced S-phase reentry. This inhibition then leads to the stabilization of cyclin E and p21cip1. Since efficient initiation of viral DNA replication requires cyclin E and cdk2, its inhibition accounts for heterogeneous viral activities in productively infected lesions.
Related Articles
Differential Regulation of P27Kip1 Expression by Mitogenic and Hypertrophic Factors
Servant, Marc J.
Coulombe, Philippe
Turgeon, Benjamin
Meloche, Sylvain
The Journal of Cell Biology
Platelet-derived growth factor-BB (PDGF-BB) acts as a full mitogen for cultured aortic smooth muscle cells (SMC), promoting DNA synthesis and cell proliferation. In contrast, angiotensin II (Ang II) induces cellular hypertrophy as a result of increased protein synthesis, but is unable to drive cells into S phase. In an effort to understand the molecular basis for this differential growth response, we have examined the downstream effects of PDGF-BB and Ang II on regulators of the cell cycle machinery in rat aortic SMC. Both PDGF-BB and Ang II were found to stimulate the accumulation of G1 cyclins with similar kinetics. In addition, little difference was observed in the expression level of their catalytic partners, Cdk4 and Cdk2. However, while both factors increased the enzymatic activity of Cdk4, only PDGF-BB stimulated Cdk2 activity in late G1 phase. The lack of activation of Cdk2 in Ang II-treated cells was causally related to the failure of Ang II to stimulate phosphorylation of the enzyme on threonine and to downregulate p27Kip1 expression. By contrast, exposure to PDGF-BB resulted in a progressive and dramatic reduction in the level of p27Kip1 protein. The time course of p27Kip1 decline was correlated with a reduced rate of synthesis and an increased rate of degradation of the protein. Importantly, the repression of p27Kip1 synthesis by PDGF-BB was associated with a marked attenuation of Kip1 gene transcription and a corresponding decrease in Kip1 mRNA accumulation. We also show that the failure of Ang II to promote S phase entry is not related to the autocrine production of transforming growth factor-β1 by aortic SMC. These results identify p27Kip1 as an important regulator of the phenotypic response of vascular SMC to mitogenic and hypertrophic stimuli.
growth factors; cell cycle; CDK inhibitors; gene expression; smooth muscle cells
Related Articles
p27Kip1 induces an accumulation of the repressor complexes of E2F and inhibits expression of the E2F-regulated genes.
Shiyanov, P
Hayes, S
Chen, N
Pestov, D G
Lau, L F
Raychaudhuri, P
Molecular Biology of the Cell
p27Kip1 is an inhibitor of the cyclin-dependent kinases and it plays an inhibitory role in the progression of cell cycle through G1 phase. To investigate the mechanism of cell cycle inhibition by p27Kip1, we constructed a cell line that inducibly expresses p27Kip1 upon addition of isopropyl-1-thio-beta-D-galactopyranoside in the culture medium. Isopropyl-1-thio-beta-D-galactopyranoside-induced expression of p27Kip1 in these cells causes a specific reduction in the expression of the E2F-regulated genes such as cyclin E, cyclin A, and dihydrofolate reductase. The reduction in the expression of these genes correlates with the p27Kip1-induced accumulation of the repressor complexes of the E2F family of factors (E2Fs). Our previous studies indicated that p21WAF1 could disrupt the interaction between cyclin/cyclin-dependent kinase 2 (cdk2) and the E2F repressor complexes E2F-p130 and E2F-p107. We show that p27Kip1, like p21WAF1, disrupts cyclin/cdk2-containing complexes of E2F-p130 leading to the accumulation of the E2F-p130 complexes, which is found in growth-arrested cells. In transient transfection assays, expression of p27Kip1 specifically inhibits transcription of a promoter containing E2F-binding sites. Mutants of p27Kip1 harboring changes in the cyclin- and cdk2-binding motifs are deficient in inhibiting transcription from the E2F sites containing reporter gene. Moreover, these mutants of p27Kip1 are also impaired in disrupting the interaction between cyclin/cdk2 and the repressor complexes of E2Fs. Taken together, these observations suggest that p27Kip1 reduces expression of the E2F-regulated genes by generating repressor complexes of E2Fs. Furthermore, the results also demonstrate that p27Kip1 inhibits expression of cyclin A and cyclin E, which are critical for progression through the G1-S phases.
Related Articles
p57KIP2 control of actin cytoskeleton dynamics is responsible for its mitochondrial pro-apoptotic effect
Kavanagh, E
Vlachos, P
Emourgeon, V
Rodhe, J
Joseph, B
Cell Death & Disease
p57 (Kip2, cyclin-dependent kinase inhibitor 1C), often found downregulated in cancer, is reported to hold tumor suppressor properties. Originally described as a cyclin-dependent kinase (cdk) inhibitor, p57KIP2 has since been shown to influence other cellular processes, beyond cell cycle regulation, including cell death and cell migration. Inhibition of cell migration by p57KIP2 is attributed to the stabilization of the actin cytoskeleton through the activation of LIM domain kinase-1 (LIMK-1). Furthermore, p57KIP2 is able to enhance mitochondrial-mediated apoptosis. Here, we report that the cell death promoting effect of p57KIP2 is linked to its effect on the actin cytoskeleton. Indeed, whereas Jasplakinolide, an actin cytoskeleton-stabilizing agent, mimicked p57KIP2's pro-apoptotic effect, destabilizing the actin cytoskeleton with cytochalsin D reversed p57KIP2's pro-apoptotic function. Conversely, LIMK-1, the enzyme mediating p57KIP2's effect on the actin cytoskeleton, was required for p57KIP2's death promoting effect. Finally, p57KIP2-mediated stabilization of the actin cytoskeleton was associated with the displacement of hexokinase-1, an inhibitor of the mitochondrial voltage-dependent anion channel, from the mitochondria, providing a possible mechanism for the promotion of the mitochondrial apoptotic cell death pathway. Altogether, our findings link together two tumor suppressor properties of p57KIP2, by showing that the promotion of cell death by p57KIP2 requires its actin cytoskeleton stabilization function.
p57KIP2; cell migration; cancer; cytoskeleton
Related Articles
Two tumor suppressors, p27Kip1 and Patched-1, collaborate to prevent medulloblastoma
Ayrault, Olivier
Zindy, Frederique
Rehg, Jerold
Sherr, Charles J.
Roussel, Martine F.
Molecular cancer research : MCR
Two cyclin-dependent kinase inhibitors, p18Ink4c and p27Kip1, are required for proper cerebellar development. Loss of either of these proteins conferred a proliferative advantage to granule neuron progenitors, although inactivation of Kip1 exerted a greater effect. Mice heterozygous for Patched1 (Ptc1+/−) that are either heterozygous or nullizygous for Kip1 developed medulloblastoma (MB) rapidly and with high penetrance. All tumors from Ptc1+/−;Kip1+/− or Ptc1+/−;Kip1−/− mice failed to express the wild type Ptc1 allele, consistent with its role as a canonical "two-hit" tumor suppressor. In contrast, expression of the wild type p27Kip1 protein was invariably maintained in MBs arising in Ptc1+/−;Kip1+/− mice, indicating that Kip1 is haploinsufficient for tumor suppression. Although MBs occurring in Ptc1+/− mice were histopathologically heterogeneous and contained intermixed regions of both rapidly proliferating and nondividing more differentiated cells, tumors that also lacked Kip1 were uniformly less differentiated, more highly proliferative, and invasive. Molecular analysis showed that the latter MBs exhibited constitutive activation of the Sonic hedgehog signaling pathway without loss of functional p53. Apart from gains or losses of single chromosomes, with gain of chromosome 6 being the most frequent, no other chromosomal anomalies were identified by spectral karyotyping, and half of the MBs so examined retained a normal karyotype. In this respect, this mouse MB model recapitulates the vast majority of human MBs that do not sustain TP53 mutations and are not aneuploid.
p27Kip1; p18Ink4c; Patched-1; medulloblastoma; cerebellum
Related Articles
Expression of constitutively active 4EBP-1 enhances p27Kip1 expression and inhibits proliferation of MCF7 breast cancer cells
Jiang, Hong
Coleman, Jennifer
Miskimins, Robin
Miskimins, W Keith
Cancer Cell International
Eukaryotic initiation factor 4E (eIF4E) is essential for cap-dependent initiation of translation. Cell proliferation is associated with increased activity of eIF4E and elevated expression of eIF4E leads to tumorigenic transformation. Many tumors express very high levels of eIF4E and this may be a critical factor in progression of the disease. In contrast, overexpression of 4EBP, an inhibitor of eIF4E, leads to cell cycle arrest and phenotypic reversion of some transformed cells.
A constitutively active form of 4EBP-1 was inducibly expressed in the human breast cancer cell line MCF7. Induction of constitutively active 4EBP-1 led to cell cycle arrest. This was not associated with a general inhibition of protein synthesis but rather with changes in specific cell cycle regulatory proteins. Cyclin D1 was downregulated while levels of the CDK inhibitor p27Kip1 were increased. The levels of cyclin E and CDK2 were unaffected but the activity of CDK2 was significantly reduced due to increased association with p27Kip1. The increase in p27Kip1 did not reflect changes in p27Kip1 mRNA or degradation rates. Rather, it was associated with enhanced synthesis of the protein, even though 4EBP-1 is expected to inhibit translation. This could be explained, at least in part, by the ability of the p27Kip1 5'-UTR to mediate cap-independent translation, which was also enhanced by expression of constitutively active 4EBP-1.
Expression of active 4EBP-1 in MCF7 leads to cell cycle arrest which is associated with downregulation of cyclin D1 and upregulation of p27Kip1. Upregulation of p27Kip1reflects increased synthesis which corresponds to enhanced cap-independent translation through the 5'-UTR of the p27Kip1 mRNA.
Related Articles
The possible role of cell cycle regulators in multistep process of HPV-associated cervical carcinoma
Bahnassy, Abeer A
Zekri, Abdel Rahman N
Saleh, Maha
Lotayef, Mohammad
Moneir, Manar
Shawki, Osama
BMC Clinical Pathology
Human papillomavirus (HPV) 16 and 18 are associated with cervical carcinogenesis through an interaction between HPV oncogenic proteins and cell cycle regulatory genes. However, the exact pathogenetic mechanisms are not determined yet.
We investigated 43 invasive squamous cell carcinoma (ISCC), 38 CIN III, 11 CINII and 18 CINI for cyclin D1, cyclin E, CDK4, p53, mdm-2, p21waf, p27, p16INK4A, Rb and Ki-67 aberrations using immunohistochemistry and molecular techniques. Twenty samples of normal cervical tissues (NCT) were taken as a control.
There was a significant increase in the expression of Ki-67, cyclin E, CDK4, p16INK4A, Rb (p= 0.003, 0.001, 0.001, 0.01) and a significant decrease in p27KIP1 from NCT to ISCC (p = 0.003). Increased cyclin D1, p21waf, p53, mdm-2 expression, homozygous deletion (HZD) and promoter methylation (PM) of the Rb were detected in CINIII and ISCC only. On univariate analysis; tumor size, differentiation, lymph node status, FIGO stage, Ki- 67, cyclin D1, p53 and p27KIP1 are significantly associated with reduced overall survival (OS) while on multivariate analysis; only FIGO stage, Ki-67, cyclin D1, p53 and p27KIP1 were significant.
1) Aberrations involving p27KIP1, cyclin E, CDK4, p16INK4A are considered early events in HPV 16 and 18-associated cervical carcinoma, whereas cyclin D1 and p53 pathway abnormalities are considered late events. 2) Immunohistochemical tests for p16INK4Aand cyclin E, could help in early diagnosis of cervical carcinoma. 3) Only FIGO stage p53, cyclin D1, p27KIP1 and Ki-67 are independent prognostic factors that might help in predicting outcome of cervical cancer patients.
Related Articles
A novel role for the cyclin-dependent kinase inhibitor p27Kip1 in angiotensin II–stimulated vascular smooth muscle cell hypertrophy
Braun-Dullaeus, Ruediger C.
Mann, Michael J.
Ziegler, Almut
von der Leyen, Heiko E.
Dzau, Victor J.
Journal of Clinical Investigation
Angiotensin II (Ang II) has been shown to stimulate either hypertrophy or hyperplasia. We postulated that the differential response of vascular smooth muscle cells (VSMCs) to Ang II is mediated by the cyclin-dependent kinase (Cdk) inhibitor p27Kip1, which is abundant in quiescent cells and drops after serum stimulation. Ang II treatment (100 nM) of quiescent VSMCs led to upregulation of the cell-cycle regulatory proteins cyclin D1, Cdk2, proliferating cell nuclear antigen, and Cdk1. p27Kip1 levels, however, remained high, and the activation of the G1-phase Cdk2 was inhibited as the cells underwent hypertrophy. Overexpression of p27Kip1 cDNA inhibited serum-stimulated [3H]thymidine incorporation compared with control-transfected cells. This cell-cycle inhibition was associated with cellular hypertrophy, as reflected by an increase in the [3H]leucine/[3H]thymidine incorporation ratio and by an increase in forward-angle light scatter during flow cytometry at 48 hours after transfection. The role of p27Kip1 in modulating the hypertrophic response of VSMCs to Ang II was further tested by antisense oligodeoxynucleotide (ODN) inhibition of p27Kip1 expression. Ang II stimulated an increase in [3H]thymidine incorporation and the percentage of S-phase cells in antisense ODN–transfected cells but not in control ODN–transfected cells. We conclude that p27Kip1 plays a role in mediating VSMC hypertrophy. Ang II stimulation of quiescent cells in which p27Kip1 levels are high results in hypertrophy but promotes hyperplasia when levels of p27Kip1 are low, as in the presence of other growth factors.
Related Articles
E Proteins and Id2 Converge on p57Kip2 To Regulate Cell Cycle in Neural Cells
Rothschild, Gerson
Zhao, Xudong
Iavarone, Antonio
Lasorella, Anna
Molecular and Cellular Biology
A precise balance between proliferation and differentiation must be maintained during neural development to obtain the correct proportion of differentiated cell types in the adult nervous system. The basic helix-loop-helix (bHLH) transcription factors known as E proteins and their natural inhibitors, the Id proteins, control the timing of differentiation and terminal exit from the cell cycle. Here we show that progression into S phase of human neuroblastoma cells is prevented by E proteins and promoted by Id2. Cyclin-dependent kinase inhibitors (CKI) have been identified as key effectors of cell cycle arrest in differentiating cells. However, p57Kip2 is the only CKI that is absolutely required for normal development. Through the use of global gene expression analysis in neuroblastoma cells engineered to acutely express the E protein E47 and Id2, we find that p57Kip2 is a target of E47. Consistent with the role of Id proteins, Id2 prevents activation of p57Kip2 expression, and the retinoblastoma tumor suppressor protein, a known Id2 inhibitor, counters this activity. The strong E47-mediated inhibition of entry into S phase is entirely reversed in cells in which expression of p57Kip2 is silenced by RNA interference. During brain development, expression of p57Kip2 is opposite that of Id2. Our findings identify p57Kip2 as a functionally relevant target recruited by bHLH transcription factors to induce cell cycle arrest in developing neuroblasts and suggest that deregulated expression of Id proteins may be an epigenetic mechanism to silence expression of this CKI in neural tumors.
Related Articles
CDK inhibitor p57Kip2 is negatively regulated by COP9 signalosome subunit 6
Chen, Bo
Zhao, Ruiying
Su, Chun-Hui
Linan, Monica
Tseng, Chieh
Phan, Liem
Fang, Lekuan
Yang, Heng-Yin
Yang, Huiling
Wang, Wenqian
Xu, Xiaoyin
Jiang, Nan
Cai, Shouliang
Jin, Feng
Yeung, Sai-Ching J.
Lee, Mong-Hong
Cell Cycle
Subunit 6 of the COP9 signalosome complex, CSN6, is known to be critical to the regulation of the MDM2-p53 axis for cell proliferation and anti-apoptosis, but its many targets remain unclear. Here we show that p57Kip2 is a target of CSN6, and that CSN6 is a negative regulator of p57Kip2. CSN6 associates with p57Kip2, and its overexpression can decrease the steady-state expression of p57Kip2; accordingly, CSN6 deficiency leads to p57Kip2 stabilization. Mechanistic studies show that CSN6 associates with p57Kip2 and Skp2, a component of the E3 ligase, which, in turn, facilitates Skp2-mediated protein ubiquitination of p57Kip2. Loss of Skp2 compromised CSN6-mediated p57Kip2 destabilization, suggesting collaboration between Skp2 and CSN6 in degradation of p57Kip2. CSN6's negative impact on p57Kip2 elevation translates into cell growth promotion, cell cycle deregulation and potentiated transformational activity. Significantly, univariate Kaplan-Meier analysis of tumor samples demonstrates that high CSN6 expression or low p57 expression is associated with poor overall survival. These data suggest that CSN6 is an important negative regulator of p57Kip2, and that overexpression of CSN6 in many types of cancer could lead to decreased expression of p57Kip2 and result in promoted cancer cell growth.
COP9; CSN6; Skp2; cell cycle; p57
Related Articles
Cyclin dependent kinase inhibitor p27Kip1 expression in normal and neoplastic cervical epithelium.
Troncone, G
Vetrani, A
de Rosa, G
Gerbasio, D
Palombini, L
Journal of Clinical Pathology
AIM: To investigate whether there is loss of the p27Kip1 protein in developing cervical cancer and whether p27Kip1 immunoreactivity has any relation to the proliferative indicator Ki-67. METHODS: The expression of p27Kip1 and Ki-67 was assessed by immunohistochemistry in serial sections from normal epithelium (13), low grade (27) and high grade (19) squamous intraepithelial lesions (LSIL, HSIL), and invasive cervical cancer (23). In the SIL cases the presence of human papillomavirus (HPV) genomic sequences was assessed by in situ hybridisation. The results were evaluated by image analysis, and reported as mean score of the percentage of p27Kip1 and of Ki-67 positive cells in each histological group. RESULTS: In general, p27Kip1 immunostaining was related to squamous differentation, and was intense in normal epithelium (47%), while it was reduced in SIL lesions as an effect of the decreased number of differentiating cells. However, decrease in the p27Kip1 expression was more evident in LSIL (36%) than in HSIL (39%); in the latter, p27Kip1 had a different intraepithelial distribution in that the staining extended to the basal cells. The average levels of p27Kip1 were similar in SIL lesions associated to low, intermediate, and high risk HPV types. Compared with normal epithelium and dysplasia, invasive cancer showed significantly lower p27Kip1 levels (23%). There was no relation between p27Kip1 and Ki-67 labelling indices in any of the histological groups examined. CONCLUSIONS: A reduction in p27Kip1 protein occurs in cervical cancer independently of the proliferative status. The changes in p27Kip1 expression may be related to the unregulated kinetics of developing cervical cancer.
Related Articles
Results 1-25 (1361244)
« Previous
Go to page number:
Next »
Remove citation from clipboard
Add citation to clipboard
This will clear all selections from your clipboard. Do you wish proceed?
Clipboard is full! Please remove an item and try again.
Contact Us
PubMed Central Canada is a service of the
Canadian Institutes of Health Research
(CIHR) working in partnership with the National Research Council's
national science library
in cooperation with the
National Center for Biotechnology Information
at the
U.S. National Library of Medicine
(NCBI/NLM). It includes content provided to the
PubMed Central International archive
by participating publishers.


text:
August 17, 2012
Auckland, New Zealand
Amelie, 15 months
I moved to Switzerland two and a half years ago. I am currently doing a part-time course in legal translation (French to English). Besides babies, I enjoy reading, films, swimming and catching up wtih friends.
Gatherings in parks and at playgrounds.
Hi, I'm Rebecca and my daughter is Amelie (15 months). We came along to the Baden meet-up and the Zurich meet-up this week with Claire and Amaia. Looking forward to future meet-ups!
Meetup members, Log in
By clicking "Sign up" or "Sign up using Facebook", you confirm that you accept our Terms of Service & Privacy Policy

subset: commoncrawl


text:
A few years ago, a group of people, looking to benefit the community, founded the Prairie Village Jazz Festival. They considered a city-wide celebration, but the price of national jazz talent quickly convinced them to scale their plans to a community event, focusing on performers from or with distinct ties to Kansas City.
|Scooped by KCStage


text:
Soros: The Life and Times of a Messianic Billionaire (Paperback)
A penniless émigré who made a fortune and became one of the great philanthropists of the twentieth century, George Soros has led a remarkable life. This biography brings forth his story in unprecedented depth, from his childhood as a Jew in occupied Budapest during World War II to his conquests on Wall Street and the establishment of his philanthropic Open Society foundations. Soros offers exclusive glimpses at an often misunderstood man, revealing a shy character whose own struggle to escape the Nazis left him with the adamant belief that people of the world are entitled to live without the fear of oppression.
Enigmatic, contradictory, and inspiring, George Soros is one of the most intriguing and globally influential men of our time. In this accomplished biography, written with Soros's cooperation, Michael T. Kaufman fully illuminates the man, his motivations, and his legacy.
About the Author
Kaufman is former deputy director of the Center for Research on Latin America and the Caribbean at York University in Toronto.
Praise For…
"A fascinating account of one of the great men of our times." –The New York Times Book Review
"Masterly . . . [Kaufman] is a likable and intelligent narrator who writes with both insight and compassion." –The Washington Post Book World
"A flinty-eyed exposition of a brilliant capitalist, devoted provocateur, and accidental humanitarian. You come away believing it is possible to be a really rich man and a really good man after all." –The New York Times
"Kaufman excels at dissecting and explaining Soros's psychological makeup." –Salon
"Kaufman's biography of Soros meets a higher truth standard than most . . . A compelling narrative notable for its candor and breadth." –The Plain Dealer
"What's memorable about Kaufman's biography is its exploration of Soros as a man who satisfied his contemplative side not by making money but by finding visionary ways to spend it." –Milwaukee Journal Sentinel
"The enthralling story of an extraordinary individual. . . . Rewarding reading on several levels–as an adventure story, as food for serious philosophical speculation, and as a peek into the world of high financial wheeling and dealing." –Fort Worth Star-Telegram
"Surprisingly even-handed. . . . A thoughtful analysis on the complex intersection of wealth and compassion."–Pittsburgh Post-Gazette
"Engaging. . . .To call Soros complex is an understatement. . . . Clearly written and thoroughly researched." –San José Mercury News
"A fascinating book about an exceedingly complicated and competitive man." –The Jewish Advocate


text:
Free personalized radio that
plays the music you love
Something In The Way
Features of This Trackr&b influences
danceable beats
prominent backup vocals
a female vocal
use of wild harmonies
the use of chordal patterning
extended harmonic patterns
an electric bass riff
subtle use of electric keys
wah-wah guitar
electric guitars
use of electric pianos
synth swoops
light synth fx
subtle use of staccato synths
These are just a few of the hundreds of attributes cataloged for this track by the Music Genome Project.show more
Similar Tracks
Don't have a Pandora account? Sign up
Already have a Pandora account? Log In


text:
Mission & Vision
We are a medical home dedicated to excellence in the comprehensive medical care of adolescents and adults through all phases of life. We deliver preventive, diagnostic, therapeutic and palliative strategies with a commitment to prompt and honest communication. We serve as a hub for efficient coordination and sharing of information among the specialized medical professionals and services our patients may need from time to time, and as an advisor in the course of complex health care decision making.Our Vision:
We are committed to the highest ethical and professional standards including dedication to transparency and avoidance of all conflict with our goal of patient advocacy, apparent and actual. We are dedicated to maintenance and expansion of our skills and credentials, service to our profession and community.
Get in touch
- 4950 S Le Jeune Rd
Coral Gables, FL 33146
One Andalusia Avenue, Suite 101
Coral Gables, FL 33134
Office Hours
Monday - Friday
Open: 8:00am
Close: 4:00pm
Available by phone 24/7
Why Choose Us?
You'll also benefit from the highest standard of medical care and personal service, including:


text:
Hassayampa Inn
122 East Gurley Street, Prescott, AZ 86301, United States of America – Great location - show map
Excellent location – rated 9.6/10! (score from 323 reviews)
Rated by guests after their stay at Hassayampa Inn.
|Room Type
|Deluxe Double Room
|Classic Queen Room
|Deluxe King Room
|King Suite
6 Reasons to Choose Hassayampa Inn
Low rates
24 properties in Prescott
Manage your bookings online
They speak 2 languages
Booking is safe
Facilities of Hassayampa Inn Great facilities! Review score, 8.5
Pets are allowed on request. Charges may apply.
- Fitness Center
Food & Drink
- Bar
- Breakfast in the Room
- Restaurant with Dining Menu
- Snack Bar
Free! WiFi is available in all areas and is free of charge.
Free! Free public parking is available on site (reservation is not needed).
- Room Service
- Vending Machine (drinks)
- Vending Machine (snacks)
- 24-Hour Front Desk
- Baggage Storage
- Laundry
- Meeting/Banquet Facilities
- Business Center
- Fax/Photocopying
- Newspapers
- Safe
- Non-smoking Rooms
- Facilities for Disabled Guests
- Family Rooms
- Heating
- Air Conditioning
Languages Spoken
- Spanish
- English
From 4:00 PM
Until 11:00 AM
Cancellation and prepayment policies vary according to room type. Please enter the dates of your stay and check what conditions apply to your preferred room.
Children and Extra Beds
All children are welcome.
Up to two children under 12 years stay free of charge when using existing beds.
One additional older child or adult is charged USD 25 per night when using existing beds.
There is no capacity for extra beds in the room.
Additional fees are not calculated automatically in the total cost and will have to be paid for separately during your stay.
Pets are allowed on request. Charges may apply.
Cards accepted at this property
Hover over the cards for more info.
1 Appealing Option:
- Holiday Inn Express Prescott has a review score of 9.4
Guests are required to show a photo ID and credit card upon check-in. Please note that all Special Requests are subject to availability and additional charges may apply.


text:
Soil & Groundwater Companies serving Tonga
Current filtering criteriaYou can remove a filter by clicking on the x
Category: Soil & Groundwater ×
Employees number: 101-1000 ×
Industry: Energy - Renewable Energy ×
Serving: Australasia ×
Serving: Tonga ×
LR Senergy
based in Newcastle upon Tyne, UNITED KINGDOM
LR Senergy is a leading provider of solutions for the grid integration of renewables, specialising in the electrical connection of renewable energy projects to grid networks; from initial concept, to design and commissioning. Operating within three core areas of Consultancy, Contracting and Technology, clients are able to call on expertise from a knowledge base that spans ...
based in Leinfelden-Echterdingen, GERMANY
thinkstep is the global leader in sustainability performance management. Our data, drawn from more than 7,000 datasets, is unrivalled in its scope and rigour, and permits precise sustainability calculations across all parts of the supply chain. Our software is robust, convenient to use and available on-demand or as part of a complex, configurable solution. Our services ...
Clarke Energy
based in Liverpool, UNITED KINGDOM
Clarke Energy is a multinational specialist in the engineering, installation and maintenance of gas-fuelled power plants. Our facilities deliver fuel efficiency and help reduce carbon emissions. Applications include combined heat and power (CHP) with natural gas, biogas, landfill gas and coal gas. We are a the largest authorised distributor and service provider for GE's ...
Ashtead Technology Ltd
based in Letchworth, UNITED KINGDOM
Ashtead Technology was formed in 1985 when Ashtead Group plc acquired SubSpek Limited, which at the time provided underwater positioning, survey and inspection equipment rental from a base in Aberdeen. In 1994 the Singapore base was opened, focussing on the offshore oil and gas industry and the subsea telecommunications cable market in Asia Pacific. The first US ...
SSI Shredding Systems, Inc.
based in Wilsonville, OREGON (USA)
SSI Shredding Systems is a leading manufacturer of industrial shredders and size reduction systems. SSI Shredding Systems custom designs and manufactures high quality 1-shaft, 2-shaft, and 4-shaft rotary shear shredders and primary waste reducers as well as pre-load transfer station and metals compactors. Our highly qualified staff provide you with complete system design ...
Presto Geosystems
based in Appleton, WISCONSIN (USA)
Presto Geosystems is a leading manufacturer of innovative, quality soil stabilization, porous pavement and construction mat products. Presto's Geoweb geocells are globally recognized for solving soil stabilization problems including slope protection, channel protection, erosion control, load support and retaining walls. Free project evaluation service available. ...
Environnement S.A
based in Poissy, FRANCE
Environnement S.A is the leading company in the field of instrumentation for the environment. Ever since its creation in 1978, Environnement S.A has been dedicated to the study, development, manufacture and supply of advanced on-line gas, particulate and water analyzers, using the most recent technologies. Environnement S.A proposes a comprehensive range of continuous ...


text:
Object Permance Boxes allow a child to practice hand eye coordination, and helps the child understand that the object still exists even though they cannot see it. This work stimulates the child's curiosity making it exciting to learn.
The set contains a wooden box with tray and a white plastic ball.
Purchasing this will earn you 28.95 rewards points!
Gross Weight: 2.20 lbs
Write an online review and share your thoughts with other viewers!
Avg. Customer Review:
Number Of Reviews: 5
10 of 19 people found the following review helpful
Well-made and toddlers love it!
Was this review helpful to you?
10 of 20 people found the following review helpful
Great for toddlers!
9 of 18 people found the following review helpful
Well made, attractive, and engaging. Holds up well to use.
This is a great item that really grabs children's attention. Fantastic for children who are exploring the idea of object permanence. As children age the item can also be used as a very basic introduction to the idea of geometric shape.
3724 of 7372 people found the following review helpful
This product looks nice and seems well made.


text:
great jam! want to see them again!
When ever I hear this band, just makes me want to dance. They are the best in Memphis!
Not because I am thee founding member.. but because we play good music... And we get down !!!
By Jackson Baker
download this issue
click here to see more »


text:
Join us in celebrating 70 years of great seafood throughout 2016.
Sign-up for our E-Club (Click Here!) to receive special offers and 70th anniversary deals!
Join us for lunch or dinner. Opens every day at 11:00 AM.
Offering an extensive Happy Hour menu with great discounts on food and drinks.
Monday through Friday, 3:30-6:30 PM
Fully renovated indoor and a brand new outdoor patio, Anthony's La Mesa is East County's premier destination restaurant.
Treat your guests to Anthony's semi-private banquets, festive holiday parties, or do something different, like host an afternoon bocce ball cocktail party.
Check out our Private Events page.


text:
2 boys and 2 girls! Mom is a Blue Razor, dad is a Blue Bully! These energetic pups are looking for their forever home! Great with kids and extremely smart these pups will bring joy to any household! First round of shots and two rounds of dewormer already given! Mom and Dad also pictured!
Jul 17 -
Greensboro :
Winston Salem - Dogs & puppies
I have for sale 5 gorgeous blue tri pitbull puppies. 3 boys and 2 girls (mostly blue ones). 1st shots given by vet today (6-21) and vet said they are good and healthy pups. UKC papers in hand ready to go. Call or text 478-235-0522
Jun 21 -
Columbus (GA) :
Montezuma - Dogs & puppies
We'll be sending text message alerts for your subscription to


text:
Minister of State
for Home Affairs's
July 19, 2016
Dr. Guruprasad Mohapatra, IAS, takes over as Chairman, Airports Authority of India
July 12, 2016
Dassault Falcon Jet names James Hurley Senior Vice President of Eastern US and Canadian Sales
July 6, 2016
Jayant Sinha takes charge as Minister of State in Civil Aviation Ministry
June 21, 2016
Embraer announces John Slattery as its new Commercial Aviation President and CEO
View All
By B.K. Pandey, Editor,
SP's Airbuz
By Group Captain R.K. Bali (Retd), MD, BAOA
By B.K. Pandey
The Asia-Pacific region accounts for 40 per cent of commercial aviation...
The state of Gujarat has been a progressive one. It is taking active steps...
Rolls-Royce is a power systems company catering to aircraft...
'Sunrise' is a traditional definition. We are looking forward to every project and we see growth.
As India's civil aviation sector works towards its potential, there are major...
India is increasingly becoming the epicenter of great innovation and it is critical to contribute...
Aggressive Plans for China, India
'Indian Market has...'
Embraer - The Game Changer in India
It is a matter of pride that "SP Guide
Publications" is working since 50 years
in the field of aerospace and defense...
read more
"I am happy to learn that SP Guide
Publications is completing 50 years
of existence."
read more
The CMD and Editor-in-Chief of SP Guide Publications briefing Mr P. Ashok Gajapathi Raju, Union Minister of Civil Aviation about the newly released magazine BizAvIndia
Rs. 600/US$180
per year
Rs. 1,320/US$325
per year
Supplement to
SP's Aviation 5/2016
A Message from
Prime Minster's office
The Hon'ble Prime Minister is happy to learn that SP Guide Publications is celebrating Golden Jubilee of its publication.
In a country like India with limited support from the industry and market, initiating 50 years ago publishing magazines relating to Army, Navy and Aviation sectors without any interruption is a commendable job on the part of SP Guide Publications...
I am happy to know that SP Guide
Publications will be completing 50 years
this year...
read more
I am glad to know that SP Guide Publications, New Delhi with its six publications mainly catering to different facets of defence and aerospace, is completing 50 years of service... read more
I am pleased to learn that SP Guide
have completed more than
52 years in bringing
out journals
on aerospace and defence...
read more


text:
Has this been fixed?
Can see there was a minor updater yesterday..
Type: Posts; User: Rhazmuz; Keyword(s):
Has this been fixed?
Can see there was a minor updater yesterday..
Must say, top notch work mate! Right up there with the best skins for FM16 :)
Arent you a charmer...
Didnt you leave the forums?
Yea he fixed it, after this was posted, and after the patch was released, which made the game crash with the mentioned skins.
Amazing how people can react like this, like small children, simply...
Sounds like sour grapes to me
Those player shirts look really, well done!
A. Laporte
I run it at 1920x1080. Besides in the OP it states: "Total player overview overhaul with 2 different layouts"
So Im guessing there is the one I quoted (with Smolnikov), which is what I am seeing...
Anyone know how to get the profile overview as in the opening post? Mine looks like the one above..
So in the first seaosn the board expects us to win the league, no way to adjust that expectation after the 16.1 patch. What are you guys' experience with this, will I be fired if I dont win the...
Top notch, merry christmas :)
Crisp mate, cant wait to see this ingame. Thx to all who helped out, what community is all about :)
Awesome work man!
Pretty impressive skin!
My only input is that the logos in the top bar are barely visible, making them more prevalent would be an improvement imo. Other than that its pretty darn slick :)
Ouff that looks smooth Tom, good job!
Are Monaco Happy to extend the loan or is there some extension clause?
How do you guys approach tough away games? Like what system do you use?
For home games I have a 4-3-1-2 system, which plays quite similar to the real life diamond, but for the games against the...
Cheers bud! :)
Ive only quickly looked over the skin, but it seems we can not see training/stat progression (the arrows next to the different attributes numbers) in the profile screen. Is it possible to add this,...
Right click the team name
Didnt work for me. I had to right click the club name and then select view club overview.
Hey Tom, great skin!
Is there any way to make the default transparency 20% instead of 40%?
Thx cachirro, worked with zippyshare :D
Nr 4, tweaked match title bar dosnt work for me, I get error #12 from sendspace when I try to download :(
Also another enquiry
Is it possible to make the title bar slightly "slimmer", as to make it fill less space on the top? Like is there a height value for it somewhere I can tinker with?


text:
No worries, always like to share products with everyone when the product is a good one!
Hahah yeah I'm fairly at home in a hammock, the vid makes it look crazy as the trees I was on gave some...
Type: Posts; User: Lepmeister
No worries, always like to share products with everyone when the product is a good one!
Hahah yeah I'm fairly at home in a hammock, the vid makes it look crazy as the trees I was on gave some...
Good job.... They will only get better from here!
Just got a new hammock and couldn't be happier. It's very comfortable and come standard with some great inclusions.
This vid is a quick run over of its features and why I liked it so much after...
I'm not going to be able to make the Sydney based one which is coming up soon but I'll stinging to get out and enjoy this...
I have found that my Superfly adds to maintaining warmth with the doors down for sure. Most noticeable with a breeze or wind. Also in the morning with dew
Nice job Shug... Ladder action was great!
I have run your method for a few years after checking out how the pro (you) did it back in the day. Still use it but have messed with a gear hammock of...
Very true!
Agree with youtube... I watched a few before getting my singer 99 and by the time i picked it up a few days later i was ready to go. Ive made heaps of stuff that i could only look at others making in...
Looks great in the red.. Well done.
Ill have to get my hands on the plans and make one myself.
Nice work! Looks the goods. You may have more stretch in a 1.1 single which will case a negitive effect in lay comfort.
I use zing it in an adjustable style. Check out the below to see how it works (start at the 1 min mark if you like).
I know! The list is growing faster than the materials are delivered :-) cheers mate
Came with the book and wooden case too!
$60 .... :-)
Thanks, im glad im into it now. I think i was always of the mind that an old singer would cost more than it did.
Cheers :-)
You may have the 99k? They seem to go for ever!
Looks just...
Question on seam sealing for next hammock tarp project...
So dont worry about the subjuct matter below, but what do you think about sealing the lower edge of tarps? Required or not?
Yeah, isn't he great.. Thats Parker our 3 mth old Golden Retiever
Thanks :-) i hadnt used one since high school some 20 years ago but a few youtube videos and i was away. Go on......
Ummmm... Shhhhhh... A search for henry shires should see you right...
Looks great - nice tips too.. I have just started on the DIY / thread injector curve and the temptation to freestyle as somthing hard to fight!
I have been watching others make some great stuff over the years on the thread injector and it was time i got in on the action.
I got myself a Singer 1951 201K
First hammock item was...
Its a gear hammock i rigged up out of my pack cover...
Full set-up shown here..
1951 201k
Im in sydney where there are a few of us who would make a group hang.
Hi Guys,
Just purchased a old singer 201K for some DIY gear fun..
Did the pattern in this thread work for the superfly size?
Great stuff... Always need to be looking out for the widow makers!
Ill be looking to do the same soon 85027
Cool, thanks for the tips. I have never seen it before, they work a treat.
Superfly as ground set up.
Whoopie inspired adjustable tarp lines
Hey guys,
Northwest Sydney, been too busy to be posting here as much as i would like. Let alone getting out and using the hammock in anything other than the backyard.
Should be getting back in...
Welcome Mate
Great stufff... That should see it right! Thanks guys
After a another great weekend away with a few mates (fellow HF members here is aust.) the urge got me and as soon as i was home to get ordering from Dutch.
Here is a quick vid of the setup i made....
That is spooky.... I'm not hanging anywhere near that!
My WB superfly always ends up getting placed in my pack. The extra size and doors just gives the peice of mind that if the wind or / rain starts up im covered.
Didnt so much change me, but sure did change what i like doing. i have meet some of my best friends on the back of this forum and hammocks!
Great location... I like the tree stand too.
Hey Guys,
I have one on the way, did HD provide tracking numbers to your when your shipped?
I'll make a set-up Vid when I get mine for future buyers :-)
Very nice, great looking spot.
Doesn't even look like the same hammock... Nice one.
I like a tree that doesn't bend when you get in your hammock. It just messes with the tension of our tarp.
Using the dropper loops from the pole of the tie outs like you have, do you think a stiff pole "hiking pole" would also work?
Nice!! I like it alot.
I have one on the way too, cant wait. Thanks for the teaser!
Looks like you enjoying the Sydney area.
Looks like a great spot. Thanks for sharing
Looking forward to mine arriving soon. Just dreaming of setting it up now!
Thanks for watching guys,
Yeah the hand drill evaded me on this my first attempt. I may try it with a bow and drill as that is easier and come back to the handdrill.
Thanks for the thoughts...
I have found in the WBBB that with my head right over near the zipper and feet in the box is the best. The ridge under your legs would have moved the UQ away from the hammock so that wouldnt have...
Great selection!


text:
Hotels near Lech Fall
Search hotels near Lech Fall in Fuessen
Trending now in Fuessen
- Most bookedVital Hotel Wiedemann
23 people looked at this hotel in the last hour
Most recent review
"The cleaning ladies started cleaning our room as we were out to breakfast. Due to our limited German they asked us to leave. They thought we were coming in for the next night. Overall nice location and service."
A Traveller, Oct 2015, GB
22 people looked at this hotel in the last hour
Most recent review
"Dinner was great and the views were fabulous. Staff were very kind and gave us the perfect information. Room and bathroom were clean as expected."
A Traveller, Jun 2016, US
22 people looked at this hotel in the last hour
Most recent review
"We required a handicapped accessible room. The room was very accessible for my wife and her wheelchair. The castles were great although a long steep hilly walk to get to the castles. (not the hotels problem, FYI only."
A Traveller, May 2014, US
20 people looked at this hotel in the last hour
Most recent review
"Nice friendly and helpful staff. Hotel is a family business, this family is just awesome. Clean room, well equipped; quiet place, comfort stay. The breakfast is serving super! Good network for web. Hotel is on a "hill top" next to the Füssen city, so you can just park your car and take a good ..."
A Traveller, May 2016, HK
17 people looked at this hotel in the last hour
Most recent review
"Very serene surroundings with a park right in front of hotel. The hotel room was very old fashioned. However, the bedding was comfortable. The bathroom amenities were from the dark ages !!! We reached late in the evening and no one is available at the reception desk. The rooms are spread over 3 ..."
A Traveller, May 2016, US
16 people looked at this hotel in the last hour
Most recent review
"Close to the train station, wonderful upgrades to a room with a wonderful view of cobblestone street. Amazing restaurant connected to the hotel. "
T, Jun 2016, US
14 people looked at this hotel in the last hour
Most recent review
"It was a nice hotel and had a very nice and friendly staff, the only thing that was hard was the hill we had to walk up to get to it! Would recommend it to everyone!"
A Traveller, Jun 2016, US
13 people looked at this hotel in the last hour
Most recent review
"Spent one night here on a 3 week European Tour. Nice hotel with comfortable beds, clean rooms, and good service."
A Traveller, May 2016, US
10 people looked at this hotel in the last hour
Most recent review
"I enjoyed my stay thoroughly: I enjoyed the walk up to the hotel and down to the old town. It's beautiful! Everything you need is within walking distance from the hotel - groceries, post office, bank and good restaurants. I walked from the central train station as well, but there are taxis th..."
A Traveller, Jun 2016, CA
- Old Kings Fuessen Design HostelNo Guest Reviews yet
Map of hotels near Lech Fall
Hotels in Fuessen near Lech Fall
Are you looking for a cheap Lech Fall hotel, a 5 star Lech Fall hotel or a family friendly Lech Fall hotel? You just landed in the best site to find the best deals and offers on the most amazing accommodations for your stay.
When you search for hotels near Lech Fall with Hotels.com, you need to first check our online map and see the distance you will be from Lech Fall, Germany.
Our maps are based on hotel search and display areas and neighborhoods of each hotel so you can see how close you are from Lech Fall and refine your search within Fuessen or Germany based on closest public transportation, restaurants and entertainment so you can easily get around the city. All the hotels details page show an option for free or paid onsite parking.
If you wish to see the hotels with the highest featuring discounts and deals near Lech Fall, simply filter by price/ average nightly rate. We recommend you filter by star rating and read our genuine guest reviews so you can get the best quality hotel with the best discount.
One of the new features on Hotels.com guest reviews is that also show reviews from Expedia for Lech Fall hotels and the TripAdvisor Lech Fall hotels reviews so you can make sure that you checking with a reliable source. See the review scores on our Fuessen hotel information pages.
Make the most out of your family vacation when you book your accommodation with Hotels.com – book your hotel near Lech Fall, Fuessen after reviewing the facilities and amenities listed for each hotel.
After booking your hotel near Lech Fall, expect to receive your reservation confirmation in the mail in less than 10 minutes. The confirmation email contains more information on all nearby attractions, local directions and weather forecast, so you can better plan the days during your trip.
After getting the best hotel rates you can still save more by winning 1 free night! That's right, book 10 nights in any hotel near Lech Fall, Fuessen and after you sign up for the Welcome Rewards program, you are eligible hotel you receive 1 night free*
The best hotel deals are here: We have Lech Fall hotel deals, Lech Fall last minute deals and offers to get you the cheapest Lech Fall hotel with our lowest price guarantee.


text:
CW500 and IT leadership skills
Read more about related IT management topics:
News : CW500 and IT leadership skills
July 29, 2016
CIO interview: Gary Steen, chief technology officer, TalkTalk
TalkTalk has overhauled security since its controversial data breach in 2015, according to CTO Gary Steen, and is investing in technology to beat its rivals on customer service
July 29, 2016
German digital bank Fidor acquired by French banking giant
Fidor CEO says merger with BPCE will allow challenger to continue its international expansion
July 28, 2016
Nordic CIO interview: Vesa Erolainen, Pöyry
Doubts were raised when Vesa Erolainen, CIO at Finnish engineering and consulting company Pöyry, outlined his IT transformation plan, but he ended up winning an award for it
July 28, 2016
Revealed: The battle for GDS – how Whitehall mandarins are trying to carve up digital strategy
Insiders say senior civil servants want to break up the Government Digital Service and return IT to its previous departmental model
In Depth : CW500 and IT leadership skills
Digital transformation gives CIOs chance to be agents of change
Whether it's the IoT, cloud, mobility, big data or analytics, there is always something new to grapple with. At Cisco Live in Las Vegas attendees found out how to keep up Continue Reading
CIO Interview: Graeme Hackland, Williams Martini Racing
Williams Martini Racing CIO Graeme Hackland discusses network infrastructure, data analytics and the future for wearable tech and augmented reality in Formula 1 motor racing Continue Reading
Make fictional expense claims a thing of the past
Some executives think that travel and entertainment (T&E) expense claims are a licence to practise their creative fiction skills Continue Reading
CIO Trends #2
Computer Weekly talks to more IT leaders than any other publication, and our reporters gather in-depth insights into the challenges facing CIOs, CTOs and CDOs. This guide offers a collection of our most popular articles for IT leaders during the second quarter of 2016, examining the effects of the UK's Brexit vote for CIOs and debating Gartner's ...
By submitting your email address, you agree to receive emails regarding relevant topic offers from TechTarget and its partners. You can withdraw your consent at any time. Contact TechTarget at 275 Grove Street, Newton, MA.
You also agree that your personal information may be transferred and processed in the United States, and that you have read and agree to the Terms of Use and the Privacy Policy.
Blog Posts : CW500 and IT leadership skills
Brexit silver linings exist for UK technology - but they are hard to find
If there's one thing you can credit the UK tech sector for, it's a generally long-term view of the future. In this time of huge Brexit uncertainty, that pragmatism is perhaps the main cause for ... Continue Reading
Where are all the women in UK technology?
Since Computer Weekly launched its list of the most influential women in UK IT in 2011, the number of initiatives championing and encouraging women in technology has grown enormously. Through ... Continue Reading
Remaining in the EU is essential if the UK wants to be a global technology leader
Here at Computer Weekly, we want what's best for the UK tech sector and the people who work there – that's why we believe that remaining in the European Union (EU) is vital at such a critical time ... Continue Reading
Opinion : CW500 and IT leadership skills
Where are the digital leaders in local government?
Brexit presents a challenge and an opportunity for digital leaders in local government – they need to work together to find the best solutions Continue Reading
Privacy Shield gets green light – for now
Privacy Shield faces inevitable legal challenges, so organisations should prepare by considering legal and practical alternatives Continue Reading
Why digital is not enough
Digital has become a buzzword, but IT leaders instead need to understand the characteristics of a 21st century digital organisation Continue Reading
Videos : CW500 and IT leadership skills
CW500 video: Don Kavanagh discusses how to architect IT for the cloud
In this video interview, Don Kavanagh, former chief architect at Home Office SIA discusses how IT architectures can be used to maximise the benefits from the cloud
CW500 video: Mark Nicholls discusses how to architect IT for the cloud
In this video interview, Mark Nicholls, head of technical solutions and security at Peabody Trust discusses how IT architectures can be used to maximise the benefits from the cloud
CW500 video: Abdul Sheikh discusses how to architect IT for the cloud
In this video interview, Abdul Sheukh, CTO at Cintra, discusses how IT architectures can be used to maximise the benefits from the cloud


text:
Prices, specials, features and availability subject to change.
Live exquisitely surrounded by luxury at the Vantage at Merrifield, located in the Mosaic District between Routes 29 and 50 on Gallows Road. The Vantage is part of an urban enclave comprising over 80 prime acres featuring exceptional architecture that will include offices, hotels, over 1,000 sq. ft. of retail, residences and other public amenities, all creating a pedestrian friendly neighborhood. Stroll amid a wide variety of name-brand and boutique stores. Take in a movie. Enjoy lunch or dinner at unique restaurants that offer a wide variety of tantalizing fares. It's not every day that you have a chance to be a part of the area's next destination location. So, take the lead and surround yourself in luxury and convenience.
Our lease terms are: 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months (Please note that lease terms may vary, are subject to change without notice, and are based on availability. Inquire with property staff for complete details).
Pet Policy: $50 pet rent. $500 non-refundable pet fee. Restrictions: Pets welcome! Breed restrictions apply. Pet deposit and monthly pet rent apply per pet. Maximum of two pets per apartment home.
Dogs Allowed. Cats Allowed. Pets Accepted. Call for service animal policy.
253 units
Contact Propertyor call
Getting an apartment initially was a little difficult given the unresponsiveness of the leasing office, but WE Read MoreLL worth the wait. Love this place!!
March 16, 2016
Once parking is resolved, I have no problem.
March 13, 2016
Pleasant staff, nice apartments. Convenient to shopping, entertainment, restaurants, and metro.
December 17, 2015
While older than many of its competitors, the structure seems to be a higher quality construction than many we Read More have seen. When amenity and public space renovations are complete, Vantage at Merrifield will be very competitive in the market. Your location is a huge selling point. The expanding restuarant/bar, movie and shopping options within the Mosaic District offer residents a very attractive and convenient urban community.
December 01, 2015
We enjoy everything about our apartment. It is a very fun area, with several outstanding dining, shopping, an Read Mored entertainment options within a 5 minute walk. Everything is safe, clean, and the people are very friendly. The staff at the Vantage were beyond helpful in the application process, move in, and the day to day needs.
October 30, 2015
Monday-Friday: 9:00am-6:00pm
Saturday: 10:00am-5:00pm
Sunday: 12:00pm-5:00pm
Kettler Management, Inc.
Don't have a Rent.com account? Create one.
Creating an account allows you to acccess your saved and contacted properties on any device. Already have an account?
Log In
We are experiencing technical difficulties at the moment. Please try again later.


text:
Live in Los Angeles, CA now....flyiing to Costa Rica for 3 weeks of unplanned backpacking through the country right after a quick trip to Las Vegas...can't wait!.
My Favorite Pages
Cabo San Lucas
Travel Map
My top travel pages
Hope your birthday is filled with all the fun you can handle!
Sending birthday greetings from India. Hope you have a great day.
You will love Costa Rica, we were in Manual Anotonio park in April...happy birthday
Happy birthday from New Jersey. Hope your trip to Costa Rica was incredible.
Happy and free in your days there, and get harmony and peaceful, and get success and strong, and get smart and attraction, and get healthy and wealthy, This is wishing you from Burma, Happy birthday....,
Happy Birthday in Advance ,Enjoy your birthday :-)
Hi, checked back in to your page and was hoping for some new adventures. Done anything fun lately?
Happy Birthday, Summer. Have a great one.


text:
'Smart' Guns Proving to Be No Quick Fix for Firearm Violence
By LESLIE WAYNE
Published: June 15, 1999
BALTIMORE— Seven years ago, a group of engineering students at Johns Hopkins University was given $2,000 and an assignment: build a ''smart'' gun, a gun that could be fired only by its owner. Using off-the-shelf equipment, they developed a prototype that, with wires hanging out of it, was neither attractive nor practical. But it worked.
''It wasn't pretty,'' recalled Stephen P. Teret, director of the Gun Policy and Research Center at the Johns Hopkins School of Public Health. ''But if three undergrads with $2,000 and no gun experience could make it, it means others could do the same, too.''
But that has not proved to be so easy.
In the decade since the first idea of a gun that could not be fired accidentally by a child or violently by a criminal who stole it, it has come to be seen as a quick fix, cited in lawsuits and in legislatures by gun-control advocates.
So far, legislation requiring that within five years all new handguns be smart guns is pending before at least eight statehouses, including those of Connecticut, New Jersey, New York and Pennsylvania. And lawsuits to hold gun makers financially responsible for not incorporating smart-gun technology have been filed by New Orleans and 17 other cities and counties.
But as the pressure for smart guns grows, it is becoming increasingly clear that they may not be the simple fix many proponents think they are. For one thing, no workable smart gun is in sight, and as with so much in the national gun debate, its development is mired in disputes, technological problems and even questions about whether smart guns are, in fact, really a smart idea.
''When it comes to smart guns, there's no great fix out there right now,'' said Joshua Horowitz, a lawyer with the Coalition to Stop Handgun Violence, a gun-control group in Washington. ''Everyone who has looked at it said there is no quick, feasible way of doing this. For people like us, there's a question of a balancing point: would smart guns increase gun sales or would they save lives? And, from the industry's perspective, no one wants to do it until they all have to do it.''
Some gun makers welcome smart guns as a marketing opportunity to improve sagging sales of handguns; other gun makers oppose them as a first step on a slippery slope toward gun control. Some gun-control advocates say smart guns could cut down on the thousands of deaths and injuries caused each year by suicide, homicide and unintentional shootings. Other gun-control advocates oppose them, saying smart guns could encourage people who might not have guns to buy them, increasing the risk of handgun violence.
While the National Rifle Association has officially remained neutral on smart-gun technology, many of its members have lobbied against it in various legislative hearings.
Still, interest in smart guns is rising.
''The first guy to invent an affordable and reliable smart gun will be a trillionaire,'' said Paul Jannuzzo, general counsel of Glock, a gun manufacturer in Smyrna, Ga. ''It's absurd to say we're hiding this technology. Why would someone be hiding it? They'd be a wealthy person.''
While the debates continue, efforts are under way in Washington to get more Government money for smart-gun research and development. The National Institute of Justice, a Federal agency, is working with Colt's Manufacturing Company to develop a prototype smart gun for law-enforcement officers. A first test model, developed with $500,000 in Government money, is expected to be delivered this summer. The goal is to cut the number of police officers killed with their own weapons by assailants, an average of 6 to 10 a year, Government statistics show.
Police groups are putting their efforts behind a Clinton Administration budget proposal to spend $4 million on smart-gun technology. Colt, along with many members of Congress, is also seeking up to $40 million in Federal grants to develop the technology, and behind-the-scenes lobbying has already begun.
''Nobody has demonstrated a model that is viable in the marketplace,'' said Steven Sliwa, chief executive of Colt.
The lineup of semiautomatic handguns made by Colt, based in Hartford, range in price at retail from about $250 for a .22-caliber target pistol to $813 for its .45-caliber Combat Commander. The smart-gun technology could add $300 to $400 to each gun, the company says.
Why all the money for research is needed is simple: It is not easy to make a smart gun. Essentially, a smart gun is a meshing of a complicated 19th-century mechanical device, the gun, with delicate and sophisticated computer engineering. Within the footprint of an existing gun -- with controlled explosions, heavy percussions and vibrations, dirty residues and high temperatures -- electronics that would have to withstand this high stress would be imbedded. It is like putting a laptop computer into a gun and then having that computer decide when the gun will work, and when it will not. This is more complicated than what electronics in car dashboards or even in missiles are asked to do.