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Articles by Kathleen Boris-Lawrie
In JoVE (1)
Other Publications (30)
- Journal of Virology
- Journal of Virology
- Journal of Virology
- Journal of Virology
- Journal of Virology
- Virology
- Virology
- Retrovirology
- AIDS Research and Human Retroviruses
- Journal of Virology
- Retrovirology
- Current HIV Research
- Nature Structural & Molecular Biology
- Journal of Virology
- Nucleic Acids Research
- Retrovirology
- Proceedings of the National Academy of Sciences of the United States of America
- Retrovirology
- Nucleic Acids Research
- RNA Biology
- The Journal of Biological Chemistry
- Nucleic Acids Research
- Retrovirology
- PLoS Pathogens
- Methods in Enzymology
- Virus Research
- Neoplasia (New York, N.Y.)
- Virology
- Methods in Molecular Biology (Clifton, N.J.)
- Journal of Molecular Biology
Articles by Kathleen Boris-Lawrie in JoVE
Выделение родственным РНК-белковых комплексов из клеток с использованием направленного на олигонуклеотиды элюция Gatikrushna Singh*1, Sarah M. Fritz*2, Arnaz Ranji2, Deepali Singh3, Kathleen Boris-Lawrie1,2 1Department of Veterinary & Biomedical Sciences, University of Minnesota, 2Department of Veterinary Biosciences, Ohio State University, 3School of Biotechnology, Gautam Buddha University
Other articles by Kathleen Boris-Lawrie on PubMed
Nuclear Interactions Are Necessary for Translational Enhancement by Spleen Necrosis Virus RU5 Journal of Virology. Apr, 2002 | Pubmed ID: 11884554 The 5' long terminal repeat of spleen necrosis virus (SNV) facilitates Rev/Rev-responsive element (RRE)-independent expression of intron-containing human immunodeficiency virus type 1 (HIV-1) gag. The SNV RU5 region, which corresponds to the 165-nucleotide 5' RNA terminus, functions in a position- and orientation-dependent manner to enhance polysome association of intron-containing HIV-1 gag RNA and also nonviral luc RNA. Evidence is mounting that association with nuclear factors during intron removal licenses mRNAs for nuclear export, efficient translation, and nonsense-mediated decay. This project addressed the relationship between the nuclear export pathway of SNV RU5-reporter RNA and translational enhancement. Results of RNA transfection experiments suggest that cytoplasmic proteins are insufficient for SNV RU5 translational enhancement of gag or luc RNA. Reporter gene assays, leptomycin B (LMB) sensitivity experiments, and RNase protection assays indicate that RU5 gag RNA accesses a nuclear export pathway that is distinct from the LMB-inhibited leucine-rich nuclear export sequence-dependent CRM1 pathway, which is used by the HIV-1 RRE. As a unique tool with which to investigate the relationship between different RNA trafficking routes and translational enhancement, SNV RU5 and Rev/RRE were combined on a single gag RNA. We observed a less-than-synergistic effect on cytoplasmic mRNA utilization. Instead, Rev/RRE diverts RU5 gag RNA to the CRM1-dependent, LMB-inhibited pathway and abrogates translational enhancement by SNV RU5. Our study is the first to show that a nuclear factor(s) directs SNV RU5-containing RNAs to a distinct export pathway that is not inhibited by LMB and programs the intron-containing transcript for translational enhancement.
RU5 of Mason-Pfizer Monkey Virus 5' Long Terminal Repeat Enhances Cytoplasmic Expression of Human Immunodeficiency Virus Type 1 Gag-pol and Nonviral Reporter RNA Journal of Virology. Oct, 2002 | Pubmed ID: 12239296 Retroviruses utilize an unspliced version of their primary transcription product as an RNA template for synthesis of viral Gag and Pol structural and enzymatic proteins. Cytoplasmic expression of the gag-pol RNA is achieved despite the lack of intron removal and the presence of a long and highly structured 5' untranslated region that inhibits efficient ribosome scanning. In this study, we have identified for the first time that the 5' long terminal repeat (LTR) of Mason-Pfizer monkey virus (MPMV) facilitates Rev/Rev-responsive element-independent expression of HIV-1 gag-pol reporter RNA. The MPMV RU5 region of the LTR is necessary and directs functional interaction with cellular posttranscriptional modulators present in human 293 and monkey COS cells but not in quail QT-6 cells and does not require any viral protein. Deletion of MPMV RU5 decreases the abundance of spliced mRNA but has little effect on cytoplasmic accumulation of unspliced gag-pol RNA despite complete elimination of detectable Gag protein production. MPMV RU5 also exerts a positive effect on the cytoplasmic expression of intronless luc RNA, and ribosomal profile analysis demonstrates that MPMV RU5 directs subcellular localization of the luc transcript to polyribosomes. Our findings have a number of similarities with those of reports on 5' terminal posttranscriptional control elements in spleen necrosis virus and human foamy virus RNA and support the model that divergent retroviruses share 5' terminal RNA elements that interact with host proteins to program retroviral RNA for productive cytoplasmic expression.
Human T-cell Lymphotropic Virus Type 1 P12I Enhances Interleukin-2 Production During T-cell Activation Journal of Virology. Oct, 2003 | Pubmed ID: 14512551 Human T-cell lymphotropic virus type 1 (HTLV-1) causes adult T-cell leukemia/lymphoma (ATLL) and a variety of lymphoproliferative disorders. The early virus-cell interactions that determine a productive infection remain unclear. However, it is well recognized that T-cell activation is required for effective retroviral integration into the host cell genome and subsequent viral replication. The HTLV-1 pX open reading frame I encoding protein, p12(I), is critical for the virus to establish persistent infection in vivo and for infection in quiescent primary lymphocytes in vitro. p12(I) localizes in the endoplasmic reticulum (ER) and cis-Golgi apparatus, increases intracellular calcium and activates nuclear factor of activated T cells (NFAT)-mediated transcription. To clarify the function of p12(I), we tested the production of IL-2 from Jurkat T cells and peripheral blood mononuclear cells (PBMC) expressing p12(I). Lentiviral vector expressed p12(I) in Jurkat T cells enhanced interleukin-2 (IL-2) production in a calcium pathway-dependent manner during T-cell receptor (TCR) stimulation. Expression of p12(I) also induced higher NFAT-mediated reporter gene activities during TCR stimulation in Jurkat T cells. In contrast, p12 expression in PBMC elicited increased IL-2 production in the presence of phorbal ester stimulation, but not during TCR stimulation. Finally, the requirement of ER localization for p12(I)-mediated NFAT activation was demonstrated and two positive regions and two negative regions in p12(I) were identified for the activation of this transcription factor by using p12(I) truncation mutants. These results are the first to indicate that HTLV-1, an etiologic agent associated with lymphoproliferative diseases, uses a conserved accessory protein to induce T-cell activation, an antecedent to efficient viral infection.
Primary Sequence and Secondary Structure Motifs in Spleen Necrosis Virus RU5 Confer Translational Utilization of Unspliced Human Immunodeficiency Virus Type 1 Reporter RNA Journal of Virology. Nov, 2003 | Pubmed ID: 14581534 The 5' long terminal repeat (LTR) of spleen necrosis virus (SNV) contains a unique posttranscriptional control element that facilitates Rev/Rev-responsive element-independent expression of unspliced human immunodeficiency virus type 1 (HIV-1) gag reporter RNA. HIV-1 Gag expression is eliminated when SNV LTR is repositioned to the 3' untranslated region or when the RU5 region is positioned in the antisense orientation. RU5 corresponds to the 5' RNA terminus, and results presented here indicate that Gag production is sustained upon introduction of transcribed spacers that reposition SNV RU5 35 to 200 nucleotides downstream. Concordant results of deletion and point mutagenesis identified two functionally redundant and synergistic motifs (designated A and C) that are necessary and sufficient for SNV RU5 activity. Enzymatic analysis of SNV RU5 RNA structure determined that A and C correspond to stem-loop structures. Quantitative RNA and protein analysis of A and C mutants revealed that the structural integrity of A and C is necessary for protein production, and loss of function correlates with little change in steady-state level, splicing efficiency, or cytoplasmic accumulation of HIV-1 gag reporter RNA. Instead, the structural mutations eliminate cytoplasmic utilization as an mRNA template for Gag protein production. Point mutations of unpaired loop-and-bulge nucleotides that maintain the structure of A eliminate activity. The results show that the unpaired UUGU loop and U-rich bulges function together and are candidate SNV RU5 binding sites for the host cell protein(s) that directs cytoplasmic utilization of unspliced HIV-1 reporter RNA.
Analysis of Synergy Between Divergent Simple Retrovirus Posttranscriptional Control Elements Virology. Dec, 2003 | Pubmed ID: 14675633 Mason-Pfizer monkey virus (MPMV) and spleen necrosis virus (SNV) are simple retroviruses that encode functionally divergent cis-acting RNA elements that use cellular proteins to facilitate nuclear export and translation of unspliced viral RNA. We tested the hypothesis that a combination of MPMV constitutive transport element (CTE) and SNV or MPMV RU5 translational enhancer on unspliced HIV-1 gag-pol reporter RNA synergistically augments Gag production. Results of transient transfection assays validate the hypothesis of synergistic augmentation in COS cells, but not 293 cells. RNA targeting experiments verified comparable responsiveness to CTE-interactive proteins tethered by RRE and RevM10Tap in COS and 293 cells. Exogeneous expression of Tap and NXT1 was necessary and sufficient to rescue Gag augmentation in 293 cells. Overexpression experiments established that CTE, but not RU5, confers the responsiveness to Tap and NXT1 and that CTE in conjunction with Tap and NXT1 conferred a 30-fold increase in translational utilization of the cytoplasmic RNA. Our results uncovered a previously unidentified role of CTE in conjunction with Tap and NXT1 in commitment to efficient cytoplasmic RNA utilization.
Long-term Infection with Retroviral Structural Gene Vector Provides Protection Against Bovine Leukemia Virus Disease in Rabbits Virology. Nov, 2004 | Pubmed ID: 15518821 Bovine leukemia virus (BLV) infection of rabbits is a tractable model system to evaluate vaccination strategies against lymphotropic retroviruses, which represent a global human health problem. We have previously developed genetically simplified BLV structural gene vector (SGV) that replicates BLV structural and enzymatic genes independently of BLV regulatory and accessory genes. Results of a 20-month study in a rabbit model demonstrated that BLV SGV induces an antiviral immunological response and lacks pathogenicity. Here, these chronically infected-BLV SGV rabbits are assessed in a proof-of-principle study of preventative vaccination against challenge with pathogenic BLV. This study commences 24 months after BLV SGV inoculation and proceeds for an additional 20 months. The previously characterized BLV SGV rabbits and age-matched control rabbits were challenged with 1 x 10(8) fetal lamb kidney/BLV producer cells. BLV SGV rabbits seroconverted upon BLV challenge, but did not progress to BLV infection nor clinical disease. By contrast, naive rabbits became infected and succumbed to lymphotropic disease. Our findings provide proof-of-principle that chronic infection with BLV SGV induces protection against BLV infection. The data indicate that SGV based on HTLV or HIV is a promising approach against lymphotropic disease by human retroviruses.
Human T Lymphotropic Virus Type-1 P30II Alters Cellular Gene Expression to Selectively Enhance Signaling Pathways That Activate T Lymphocytes Retrovirology. Nov, 2004 | Pubmed ID: 15560845 Human T-lymphotropic virus type-1 (HTLV-1) is a deltaretrovirus that causes adult T-cell leukemia/lymphoma and is implicated in a variety of lymphocyte-mediated disorders. HTLV-1 contains both regulatory and accessory genes in four pX open reading frames. pX ORF-II encodes two proteins, p13II and p30II, which are incompletely defined in the virus life cycle or HTLV-1 pathogenesis. Proviral clones of the virus with pX ORF-II mutations diminish the ability of the virus to maintain viral loads in vivo. Exogenous expression of p30II differentially modulates CREB and Tax-responsive element-mediated transcription through its interaction with CREB-binding protein/p300 and represses tax/rex RNA nuclear export.
Human T Lymphotropic Virus Type 1 Accessory Protein P12I Modulates Calcium-mediated Cellular Gene Expression and Enhances P300 Expression in T Lymphocytes AIDS Research and Human Retroviruses. Apr, 2005 | Pubmed ID: 15943569 Human T-lymphotropic virus type 1 (HTLV-1) is the etiologic agent of adult T cell leukemia/lymphoma (ATLL), an aggressive CD4+ T lymphocyte malignancy. Activation of T lymphocytes is required for effective retroviral integration into the host cell genome and subsequent viral replication, but the molecular mechanisms involved in HTLV-1-mediated T cell activation remain unclear. HTLV-1 encodes various accessory proteins such as p12I, which has been demonstrated to be critical for HTLV-1 infectivity in vivo in rabbits and in vitro in quiescent primary human T lymphocytes. This hydrophobic protein localizes in the endoplasmic reticulum, increases intracellular calcium, and activates nuclear factor of activated T cell-mediated transcription. To further elucidate the role of p12I in regulation of cellular gene expression, we performed gene array analysis on stable p12I-expressing Jurkat T cells, using Affymetrix U133A arrays. Our data indicate that p12I altered the expression of genes associated with a network of interrelated pathways including T cell signaling, cell proliferation, and apoptosis. Expression of several calcium-regulated genes was found to be altered by p12I, consistent with known properties of the viral protein. Gene array findings were confirmed by semiquantitative RT-PCR in Jurkat T cells and primary CD4+ T lymphocytes. Furthermore, dose-dependent expression of p12I in Jurkat T cells resulted in significant increases in p300 and p300-dependent transcription. This is the first report of a viral protein influencing the transcription of p300, a rate-limiting coadapter critical in HTLV-1-mediated T cell activation. Collectively, our data strongly indicate that HTLV-1 p12I modulates cellular gene expression patterns to hasten the activation of T lymphocytes and thereby promote efficient viral infection.
Human T-cell Leukemia Virus Open Reading Frame II Encodes a Posttranscriptional Repressor That is Recruited at the Level of Transcription Journal of Virology. Jan, 2006 | Pubmed ID: 16352542 Human T-cell leukemia virus (HTLV) infection is a chronic, lifelong infection that is associated with the development of leukemia and neurological disease after a long latency period, and the mechanism by which the virus is able to evade host immune surveillance is elusive. Besides the structural and enzymatic proteins, HTLV encodes regulatory (Tax and Rex) and accessory (open reading frame I [ORF I] and ORF II) proteins. Tax activates viral and cellular transcription and promotes T-cell growth and malignant transformation. Rex acts posttranscriptionally to facilitate cytoplasmic expression of incompletely spliced viral mRNAs. Recently, we reported that the accessory gene products of HTLV-1 and HTLV-2 ORF II (p30II and p28II, respectively) are able to restrict viral replication. These proteins act as negative regulators of both Tax and Rex by binding to and retaining their mRNA in the nucleus, leading to reduced protein expression and virion production. Here, we show that p28II is recruited to the viral promoter in a Tax-dependent manner. After recruitment to the promoter, p28II or p30II then travels with the transcription elongation machinery until its target mRNA is synthesized. Experiments artificially directing these proteins to the promoter indicate that p28II, unlike HTLV-1 p30II, displays no transcriptional activity. Furthermore, the tethering of p28II directly to tax/rex mRNA resulted in repression of Tax function, which could be attributed to the ability of p28II to block TAP/p15-mediated enhancement of Tax expression. p28II-mediated reduction of viral replication in infected cells may permit survival of the cells by allowing escape from immune recognition, which is consistent with the critical role of HTLV accessory proteins in viral persistence in vivo.
Coordinate Enhancement of Transgene Transcription and Translation in a Lentiviral Vector Retrovirology. Feb, 2006 | Pubmed ID: 16480517 Coordinate enhancement of transgene transcription and translation would be a potent approach to significantly improve protein output in a broad array of viral vectors and nonviral expression systems. Many vector transgenes are complementary DNA (cDNA). The lack of splicing can significantly reduce the efficiency of their translation. Some retroviruses contain a 5' terminal post-transcriptional control element (PCE) that facilitates translation of unspliced mRNA. Here we evaluated the potential for spleen necrosis virus PCE to stimulate protein production from HIV-1 based lentiviral vector by: 1) improving translation of the internal transgene transcript; and 2) functionally synergizing with a transcriptional enhancer to achieve coordinate increases in RNA synthesis and translation.
Retrovirus Translation Initiation: Issues and Hypotheses Derived from Study of HIV-1 Current HIV Research. Apr, 2006 | Pubmed ID: 16611053 Human immunodeficiency virus type 1 (HIV-1) has a small, multifunctional genome that encodes a relatively large and complex proteome. The virus has adopted specialized post-transcriptional control mechanisms to maximize its coding capacity while economically maintaining the information stored in cis-acting replication sequences. The conserved features of the 5' untranslated region of all viral transcripts suggest they are poor substrates for cap-dependent ribosome scanning and provide a compelling rationale for internal initiation of translation. This article summarizes key experimental results of studies that have evaluated HIV-1 translation initiation. A model is discussed in which cap-dependent and cap-independent initiation mechanisms of HIV-1 co-exist to ensure viral protein production in the context of 1) structured replication motifs that inhibit ribosome scanning, and 2) alterations in host translation machinery in response to HIV-1 infection or other cellular stresses. We discuss key issues that remain to be understood and suggest parameters to validate internal initiation activity in HIV-1 and other retroviruses.
RNA Helicase A is Necessary for Translation of Selected Messenger RNAs Nature Structural & Molecular Biology. Jun, 2006 | Pubmed ID: 16680162 RNA helicase A (RHA) is a highly conserved DEAD-box protein that activates transcription, modulates RNA splicing and binds the nuclear pore complex. The life cycle of typical mRNA involves RNA processing and translation after ribosome scanning of a relatively unstructured 5' untranslated region (UTR). The precursor RNAs of retroviruses and selected cellular genes harbor a complex 5' UTR and use a yet-to-be-identified host post-transcriptional effector to stimulate efficient translation. Here we show that RHA recognizes a structured 5'-terminal post-transcriptional control element (PCE) of a retrovirus and the JUND growth-control gene. RHA interacts with PCE RNA in the nucleus and cytoplasm, facilitates polyribosome association and is necessary for its efficient translation. Our results reveal a previously unidentified role for RHA in translation and implicate RHA as an integrative effector in the continuum of gene expression from transcription to translation.
Tertiary Structural and Functional Analyses of a Viroid RNA Motif by Isostericity Matrix and Mutagenesis Reveal Its Essential Role in Replication Journal of Virology. Sep, 2006 | Pubmed ID: 16912306 RNA-templated RNA replication is essential for viral or viroid infection, as well as for regulation of cellular gene expression. Specific RNA motifs likely regulate various aspects of this replication. Viroids of the Pospiviroidae family, as represented by the Potato spindle tuber viroid (PSTVd), replicate in the nucleus by utilizing DNA-dependent RNA polymerase II. We investigated the role of the loop E (sarcin/ricin) motif of the PSTVd genomic RNA in replication. A tertiary-structural model of this motif, inferred by comparative sequence analysis and comparison with nuclear magnetic resonance and X-ray crystal structures of loop E motifs in other RNAs, is presented in which core non-Watson-Crick base pairs are precisely specified. Isostericity matrix analysis of these base pairs showed that the model accounts for the reported natural sequence variations and viable experimental mutations in loop E motifs of PSTVd and other viroids. Furthermore, isostericity matrix analysis allowed us to design disruptive, as well as compensatory, mutations of PSTVd loop E. Functional analyses of such mutants by in vitro and in vivo experiments demonstrated that loop E structural integrity is crucial for replication, specifically during transcription. Our results suggest that the PSTVd loop E motif exists and functions in vivo and provide loss-of-function genetic evidence for the essential role of a viroid RNA three-dimensional motif in rolling-circle replication. The use of isostericity matrix analysis of non-Watson-Crick base pairing to rationalize mutagenesis of tertiary motifs and systematic in vitro and in vivo functional assays of mutants offers a novel, comprehensive approach to elucidate the tertiary-structure-function relationships for RNA motifs of general biological significance.
RNA Helicase A Interacts with Divergent Lymphotropic Retroviruses and Promotes Translation of Human T-cell Leukemia Virus Type 1 Nucleic Acids Research. 2007 | Pubmed ID: 17426138 The 5' untranslated region (UTR) of retroviruses contain structured replication motifs that impose barriers to efficient ribosome scanning. Two RNA structural motifs that facilitate efficient translation initiation despite a complex 5' UTR are internal ribosome entry site (IRES) and 5' proximal post-transcriptional control element (PCE). Here, stringent RNA and protein analyses determined the 5' UTR of spleen necrosis virus (SNV), reticuloendotheliosis virus A (REV-A) and human T-cell leukemia virus type 1 (HTLV-1) exhibit PCE activity, but not IRES activity. Assessment of SNV translation initiation in the natural context of the provirus determined that SNV is reliant on a cap-dependent initiation mechanism. Experiments with siRNAs identified that REV-A and HTLV-1 PCE modulate post-transcriptional gene expression through interaction with host RNA helicase A (RHA). Analysis of hybrid SNV/HTLV-1 proviruses determined SNV PCE facilitates Rex/Rex responsive element-independent Gag production and interaction with RHA is necessary. Ribosomal profile analyses determined that RHA is necessary for polysome association of HTLV-1 gag and provide direct evidence that RHA is necessary for efficient HTLV-1 replication. We conclude that PCE/RHA is an important translation regulatory axis of multiple lymphotropic retroviruses. We speculate divergent retroviruses have evolved a convergent RNA-protein interaction to modulate translation of their highly structured mRNA.
HIV-1 Tat RNA Silencing Suppressor Activity is Conserved Across Kingdoms and Counteracts Translational Repression of HIV-1 Proceedings of the National Academy of Sciences of the United States of America. Jan, 2009 | Pubmed ID: 19122141 The RNA silencing pathway is an intracellular innate response to virus infections and retro-transposons. Many plant viruses counter this host restriction by RNA silencing suppressor (RSS) activity of a double-stranded RNA-binding protein, e.g., tomato bushy stunt virus P19. Here, we demonstrate P19 and HIV-1 Tat function across the plant and animal kingdoms and suppress a common step in RNA silencing that is downstream of small RNA maturation. Our experiments reveal that RNA silencing in HIV-1 infected human cells severely attenuates the translational output of the unspliced HIV-1 gag mRNA, and possibly all HIV-1 transcripts. The attenuation in gag mRNA translation is exacerbated by K51A substitution in the Tat double-stranded RNA-binding domain. Tat, plant virus RSS, or Dicer downregulation rescues robust gag translation and bolsters HIV-1 virion production. The reversal of HIV-1 translation repression by plant RSS supports the recent finding in Arabidopsis that plant miRNAs operate by translational inhibition. Our results identify common features between RNA silencing suppression of plant and animal viruses. We suggest that RNA silencing-mediated translation repression plays a strategic role in determining the viral set-point in a newly HIV-1-infected patient.
Mechanisms Employed by Retroviruses to Exploit Host Factors for Translational Control of a Complicated Proteome Retrovirology. Jan, 2009 | Pubmed ID: 19166625 Retroviruses have evolved multiple strategies to direct the synthesis of a complex proteome from a single primary transcript. Their mechanisms are modulated by a breadth of virus-host interactions, which are of significant fundamental interest because they ultimately affect the efficiency of virus replication and disease pathogenesis. Motifs located within the untranslated region (UTR) of the retroviral RNA have established roles in transcriptional trans-activation, RNA packaging, and genome reverse transcription; and a growing literature has revealed a necessary role of the UTR in modulating the efficiency of viral protein synthesis. Examples include a 5' UTR post-transcriptional control element (PCE), present in at least eight retroviruses, that interacts with cellular RNA helicase A to facilitate cap-dependent polyribosome association; and 3' UTR constitutive transport element (CTE) of Mason-Pfizer monkey virus that interacts with Tap/NXF1 and SR protein 9G8 to facilitate RNA export and translational utilization. By contrast, nuclear protein hnRNP E1 negatively modulates HIV-1 Gag, Env, and Rev protein synthesis. Alternative initiation strategies by ribosomal frameshifting and leaky scanning enable polycistronic translation of the cap-dependent viral transcript. Other studies posit cap-independent translation initiation by internal ribosome entry at structural features of the 5' UTR of selected retroviruses. The retroviral armamentarium also commands mechanisms to counter cellular post-transcriptional innate defenses, including protein kinase R, 2',5'-oligoadenylate synthetase and the small RNA pathway. This review will discuss recent and historically-recognized insights into retrovirus translational control. The expanding knowledge of retroviral post-transcriptional control is vital to understanding the biology of the retroviral proteome. In a broad perspective, each new insight offers a prospective target for antiviral therapy and strategic improvement of gene transfer vectors.
RNA Helicase A Modulates Translation of HIV-1 and Infectivity of Progeny Virions Nucleic Acids Research. Mar, 2010 | Pubmed ID: 20007598 Retroviruses rely on host RNA-binding proteins to modulate various steps in their replication. Previously several animal retroviruses were determined to mediate Dhx9/RNA helicase A (RHA) interaction with a 5' terminal post-transcriptional control element (PCE) for efficient translation. Herein PCE reporter assays determined HTLV-1 and HIV-1 RU5 confer orientation-dependent PCE activity. The effect of Dhx9/RHA down-regulation and rescue with siRNA-resistant RHA on expression of HIV-1(NL4-3) provirus determined that RHA is necessary for efficient HIV-1 RNA translation and requires ATPase-dependent helicase function. Quantitative analysis determined HIV-1 RNA steady-state and cytoplasmic accumulation were not reduced; rather the translational activity of viral RNA was reduced. Western blotting determined that RHA-deficient virions assemble with Lys-tRNA synthetase, exhibit processed reverse transcriptase and contain similar level of viral RNA, but they are poorly infectious on primary lymphocytes and HeLa cells. The results demonstrate RHA is an important host factor within the virus-producer cell and within the viral particle. The identification of RHA-dependent PCE activity in cellular junD RNA and in six of seven genera of Retroviridae suggests conservation of this translational control mechanism among vertebrates, and convergent evolution of Retroviridae to utilize this host mechanism.
RNA Helicases: Emerging Roles in Viral Replication and the Host Innate Response RNA Biology. Nov-Dec, 2010 | Pubmed ID: 21173576 RNA helicases serve multiple roles at the virus-host interface. In some situations, RNA helicases are essential host factors to promote viral replication; however, in other cases they serve as a cellular sensor to trigger the antiviral state in response to viral infection. All family members share the conserved ATP-dependent catalytic core linked to different substrate recognition and protein-protein interaction domains. These flanking domains can be shuffled between different helicases to achieve functional diversity. This review summarizes recent studies, which have revealed two types of activity by RNA helicases. First, RNA helicases are catalysts of progressive RNA-protein rearrangements that begin at gene transcription and culminate in mRNA translation. Second, RNA helicases can act as a scaffold for alternative protein-protein interactions that can defeat the antiviral state. The mounting fundamental understanding of RNA helicases is being used to develop selective and efficacious drugs against human and animal pathogens. The analysis of RNA helicases in virus model systems continues to provide insights into virology, cell biology and immunology, and has provided fresh perspective to continue unraveling the complexity of virus-host interactions.
Features of Double-stranded RNA-binding Domains of RNA Helicase A Are Necessary for Selective Recognition and Translation of Complex MRNAs The Journal of Biological Chemistry. Feb, 2011 | Pubmed ID: 21123178 The DExH protein RNA helicase A (RHA) plays numerous roles in cell physiology, and post-transcriptional activation of gene expression is a major role among them. RHA selectively activates translation of complex cellular and retroviral mRNAs. Although RHA requires interaction with structural features of the 5'-UTR of these target mRNAs, the molecular basis of their translation activation by RHA is poorly understood. RHA contains a conserved ATPase-dependent helicase core that is flanked by two α-β-β-β-α double-stranded RNA-binding domains at the N terminus and repeated arginine-glycine residues at the C terminus. The individual recombinant N-terminal, central helicase, and C-terminal domains were evaluated for their ability to specifically interact with cognate RNAs by in vitro biochemical measurements and mRNA translation assays in cells. The results demonstrate that N-terminal residues confer selective interaction with retroviral and junD target RNAs. Conserved lysine residues in the distal α-helix of the double-stranded RNA-binding domains are necessary to engage structural features of retroviral and junD 5'-UTRs. Exogenous expression of the N terminus coprecipitates junD mRNA and inhibits the translation activity of endogenous RHA. The results indicate that the molecular basis for the activation of translation by RHA is recognition of target mRNA by the N-terminal domain that tethers the ATP-dependent helicase for rearrangement of the complex 5'-UTR.
Evidence That Lin28 Stimulates Translation by Recruiting RNA Helicase A to Polysomes Nucleic Acids Research. May, 2011 | Pubmed ID: 21247876 The stem cell protein Lin28 functions to inhibit the biogenesis of a group of miRNAs but also stimulates the expression of a subset of mRNAs at the post-transcriptional level, the underlying mechanism of which is not yet understood. Here we report the characterization of the molecular interplay between Lin28 and RNA helicase A (RHA) known to play an important role in remodeling ribonucleoprotein particles during translation. We show that reducing Lin28 expression results in decreased RHA association with polysomes while increasing Lin28 expression leads to elevated RHA association. Further, the carboxyl terminus of Lin28 is necessary for interaction with both the amino and carboxyl termini of RHA. Importantly, a carboxyl terminal deletion mutant of Lin28 that retains RNA-binding activity fails to interact with RHA and exhibits dominant-negative effects on Lin28-dependent stimulation of translation. Taken together, these results lead us to suggest that Lin28 may stimulate translation by actively recruiting RHA to polysomes.
Tat RNA Silencing Suppressor Activity Contributes to Perturbation of Lymphocyte MiRNA by HIV-1 Retrovirology. May, 2011 | Pubmed ID: 21569500 MicroRNA (miRNA)-mediated RNA silencing is integral to virtually every cellular process including cell cycle progression and response to virus infection. The interplay between RNA silencing and HIV-1 is multifaceted, and accumulating evidence posits a strike-counterstrike interface that alters the cellular environment to favor virus replication. For instance, miRNA-mediated RNA silencing of HIV-1 translation is antagonized by HIV-1 Tat RNA silencing suppressor activity. The activity of HIV-1 accessory proteins Vpr/Vif delays cell cycle progression, which is a process prominently modulated by miRNA. The expression profile of cellular miRNA is altered by HIV-1 infection in both cultured cells and clinical samples. The open question stands of what, if any, is the contribution of Tat RNA silencing suppressor activity or Vpr/Vif activity to the perturbation of cellular miRNA by HIV-1.
Thriving Under Stress: Selective Translation of HIV-1 Structural Protein MRNA During Vpr-mediated Impairment of EIF4E Translation Activity PLoS Pathogens. 2012 | Pubmed ID: 22457629 Translation is a regulated process and is pivotal to proper cell growth and homeostasis. All retroviruses rely on the host translational machinery for viral protein synthesis and thus may be susceptible to its perturbation in response to stress, co-infection, and/or cell cycle arrest. HIV-1 infection arrests the cell cycle in the G2/M phase, potentially disrupting the regulation of host cell translation. In this study, we present evidence that HIV-1 infection downregulates translation in lymphocytes, attributable to the cell cycle arrest induced by the HIV-1 accessory protein Vpr. The molecular basis of the translation suppression is reduced accumulation of the active form of the translation initiation factor 4E (eIF4E). However, synthesis of viral structural proteins is sustained despite the general suppression of protein production. HIV-1 mRNA translation is sustained due to the distinct composition of the HIV-1 ribonucleoprotein complexes. RNA-coimmunoprecipitation assays determined that the HIV-1 unspliced and singly spliced transcripts are predominantly associated with nuclear cap binding protein 80 (CBP80) in contrast to completely-spliced viral and cellular mRNAs that are associated with eIF4E. The active translation of the nuclear cap binding complex (CBC)-bound viral mRNAs is demonstrated by ribosomal RNA profile analyses. Thus, our findings have uncovered that the maintenance of CBC association is a novel mechanism used by HIV-1 to bypass downregulation of eIF4E activity and sustain viral protein synthesis. We speculate that a subset of CBP80-bound cellular mRNAs contribute to recovery from significant cellular stress, including human retrovirus infection.
Determination of Host RNA Helicases Activity in Viral Replication Methods in Enzymology. 2012 | Pubmed ID: 22713331 RNA helicases are encoded by all eukaryotic and prokaryotic cells and a minority of viruses. Activity of RNA helicases is necessary for all steps in the expression of cells and viruses and the host innate response to virus infection. Their vast functional repertoire is attributable to the core ATP-dependent helicase domain in conjunction with flanking domains that are interchangeable and engage viral and cellular cofactors. Here, we address the important issue of host RNA helicases that are necessary for replication of a virus. This chapter covers approaches to identification and characterization of candidate helicases and methods to define the biochemical and biophysical parameters of specificity and functional activity of the enzymes. We discuss the context of cellular RNA helicase activity and virion-associated RNA helicases. The methodology and choice of controls fosters the assessment of the virologic scope of RNA helicases across divergent cell lineages and viral replication cycles.
Cellular RNA Helicases and HIV-1: Insights from Genome-wide, Proteomic, and Molecular Studies Virus Research. Feb, 2013 | Pubmed ID: 22814432 RNA helicases are ubiquitous in plants and animals and function in many cellular processes. Retroviruses, such as human immunodeficiency virus (HIV-1), encode no RNA helicases in their genomes and utilize host cellular RNA helicases at various stages of their life cycle. Here, we briefly summarize the roles RNA helicases play in HIV-1 replication that have been identified recently, in part, through genome-wide screenings, proteomics, and molecular studies. Some of these helicases augment virus propagation while others apparently participate in antiviral defenses against viral replication.
Stress-induced Isoforms of MDM2 and MDM4 Correlate with High-grade Disease and an Altered Splicing Network in Pediatric Rhabdomyosarcoma Neoplasia (New York, N.Y.). Sep, 2013 | Pubmed ID: 24027430 Pediatric rhabdomyosarcoma (RMS) is a morphologically and genetically heterogeneous malignancy commonly classified into three histologic subtypes, namely, alveolar, embryonal, and anaplastic. An issue that continues to challenge effective RMS patient prognosis is the dearth of molecular markers predictive of disease stage irrespective of tumor subtype. Our study involving a panel of 70 RMS tumors has identified specific alternative splice variants of the oncogenes Murine Double Minute 2 (MDM2) and MDM4 as potential biomarkers for RMS. Our results have demonstrated the strong association of genotoxic-stress inducible splice forms MDM2-ALT1 (91.6% Intergroup Rhabdomyosarcoma Study Group stage 4 tumors) and MDM4-ALT2 (90.9% MDM4-ALT2-positive T2 stage tumors) with high-risk metastatic RMS. Moreover, MDM2-ALT1-positive metastatic tumors belonged to both the alveolar (50%) and embryonal (41.6%) subtypes, making this the first known molecular marker for high-grade metastatic disease across the most common RMS subtypes. Furthermore, our results show that MDM2-ALT1 expression can function by directly contribute to metastatic behavior and promote the invasion of RMS cells through a matrigel-coated membrane. Additionally, expression of both MDM2-ALT1 and MDM4-ALT2 increased anchorage-independent cell-growth in soft agar assays. Intriguingly, we observed a unique coordination in the splicing of MDM2-ALT1 and MDM4-ALT2 in approximately 24% of tumor samples in a manner similar to genotoxic stress response in cell lines. To further explore splicing network alterations with possible relevance to RMS disease, we used an exon microarray approach to examine stress-inducible splicing in an RMS cell line (Rh30) and observed striking parallels between stress-responsive alternative splicing and constitutive splicing in RMS tumors.
HIV-1 and Two Avian Retroviral 5' Untranslated Regions Bind Orthologous Human and Chicken RNA Binding Proteins Virology. Dec, 2015 | Pubmed ID: 26584240 Essential host cofactors in retrovirus replication bind cis-acting sequences in the 5'untranslated region (UTR). Although host RBPs are crucial to all aspects of virus biology, elucidating their roles in replication remains a challenge to the field. Here RNA affinity-coupled-proteomics generated a comprehensive, unbiased inventory of human and avian RNA binding proteins (RBPs) co-isolating with 5'UTRs of HIV-1, spleen necrosis virus and Rous sarcoma virus. Applying stringent biochemical and statistical criteria, we identified 185 RBP; 122 were previously implicated in retrovirus biology and 63 are new to the 5'UTR proteome. RNA electrophoretic mobility assays investigated paralogs present in the common ancestor of vertebrates and one hnRNP was identified as a central node to the biological process-anchored networks of HIV-1, SNV, and RSV 5' UTR-proteomes. This comprehensive view of the host constituents of retroviral RNPs is broadly applicable to investigation of viral replication and antiviral response in both human and avian cell lineages.
Isolation of Cognate Cellular and Viral Ribonucleoprotein Complexes of HIV-1 RNA Applicable to Proteomic Discovery and Molecular Investigations Methods in Molecular Biology (Clifton, N.J.). 2016 | Pubmed ID: 26714709 All decisions affecting the life cycle of human immunodeficiency virus (HIV-1) RNA are executed by ribonucleoprotein complexes (RNPs). HIV-1 RNA cycles through a progression of host RNPs composed of RNA-binding proteins regulating all stages of synthesis, processing, nuclear transport, translation, decay, and co-localization with assembling virions. RNA affinity chromatography is a versatile method to identify RNA-binding proteins to investigate the molecular basis of viral and cellular posttranscriptional control of gene expression. The bait is a HIV-1 RNA motif immobilized on a solid support, typically magnetic or Sepharose beads. The prey is pre-formed RNPs admixed in lysate from cells or concentrated virus particles. The methodology distinguishes high-affinity RNA-protein interactions from low-affinity complexes by increases in ionic strength during progressive elution cycles. Here, we describe RNA affinity chromatography of the 5' untranslated region of HIV-1, obtaining mixtures of high-affinity RNA binding proteins suitable for mass spectrometry and proteome identification.
DHX9/RHA Binding to the PBS-Segment of the Genomic RNA During HIV-1 Assembly Bolsters Virion Infectivity Journal of Molecular Biology. Jun, 2016 | Pubmed ID: 27107641 Cellular RNA-binding proteins incorporated into virions during human immunodeficiency virus type 1 (HIV-1) assembly promote the replication efficiency of progeny virions. Despite its critical role in bolstering virion infectivity, the molecular basis for the incorporation of DHX9/RNA helicase A (RHA) to virions remains unclear. Here, cell-based experiments demonstrate that the truncation of segments of the HIV-1 5'-untranslated region (5'-UTR) distinct from the core encapsidation sequence eliminated virion incorporation of RHA, indicating that RHA recruitment is mediated by specific interactions with the HIV-1 5'-UTR. In agreement with biological data, isothermal titration calorimetry determined that the dimer conformation of the 5'-UTR binds one RHA molecule per RNA strand, and the interaction is independent of nucleocapsid protein binding. NMR spectra employing a deuterium-labeling approach enabled resolution of the dimeric 5'-UTR in complex with the RHA N-terminal domain. The structure of the large molecular mass complex was dependent on RHA binding to a double-stranded region of the primer binding site (PBS)-segment of the 5'-UTR. A single A-to-C substitution was sufficient to disrupt biophysical conformation and attenuate virion infectivity in cell-based assays. Taken together, our studies demonstrate the structural basis for HIV-1 genomic RNA to recruit beneficial cellular cofactor to virions. The support of progeny virion infectivity by RHA is attributable to structure-dependent binding at the PBS-segment of the HIV-1 5'-UTR during virus assembly.


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6489 Main Street
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Phone: 804.693.1217
Fax: 804.824-2442
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The Chesapeake Bay Preservation Act (act), section 10.1-2100 et. seq. and section 15.2-2283 of the Code of Virginia, recognizes that healthy state and local economies are integrally related to each other and the environmental health of the Chesapeake Bay. The purpose of this chapter is to control and regulate runoff at the source to protect against and minimize pollution and deposition of sediment in wetlands, streams and lakes in Gloucester County which are tributaries of the Chesapeake Bay. This chapter is intended to assist in protection of the Chesapeake Bay and its tributaries from nonpoint source pollution from land uses or appurtenances within the Chesapeake Bay drainage area. Regulations in this chapter are necessary for: (1) protection of existing high quality state waters and restoration of all other state waters to a condition or quality that will permit all reasonable public uses and will support the propagation and growth of all aquatic life, including game fish, which might reasonably be expected to inhabit them; (2) safeguarding the clean waters of the commonwealth from pollution; (3) prevention of any increase in pollution; (4) reduction of existing pollution; and (5) promotion of water resource conservation in order to provide for the health, safety and welfare of present and future citizens of the commonwealth.
This chapter establishes criteria used by Gloucester County in granting, denying or modifying requests to subdivide or develop land in Chesapeake Bay Preservation Areas.
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subset: commoncrawl


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GALVESTON, TX --It's been a long time coming, but Galveston's Municipal Skate Park is finally open, and hundreds of enthusiasts are already big fans!The city officially broke ground on the skate park in January of this year, but community support for the project originated more than seven years ago. Following the development of an ordinance banning skateboarding, local enthusiasts organized and began a grassroots campaign to raise money for a park. Eight years later, the skaters finally have a place to call home.
Grindline, an internationally known firm that is dedicated to state-of-the-art skate park design and construction, was awarded the $285,439 contract in December 2007. Park funding ultimately included many cash and in-kind donations, in addition to the city's donation of land for the site and Community Development Block Grant money.
The skate park is located in Galveston's Menard Park at 2119 27th Street. It's open from 6am to 10pm daily. The park is free.
Skaters are warned they skate at their own risk. Riders are required to wear a helmet and pads. No bicycles or roller blades are allowed.


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Actress Anna Camp has seemingly moved on from her failed marriage by striking up a new romance with her Pitch Perfect co-star Skylar Astin, according to a new report.
Actor Michael Mosley filed for divorce from Camp in April (13), citing 1 January (13) as the date of the couple's separation and blaming the split on irreconcilable differences.
Camp has posted pictures of herself with Astin on her Instagram.com page in recent weeks, and the movie co-stars stepped out together at the 24 Hour Plays event in California last weekend (22Jun13) - and now editors at UsMagazine.com claim the actress is dating the actor.
A source tells the website, "They left together. Apparently backstage Skylar was talking about how he loved her from the first day they met."
Camp married Mosley in 2010.


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April 13, 2017
Dear F2L™ VIP Guest:
I am delighted you are joining me on June 3rd for the 7th Free to Laugh: Laugh to Free™ Phoenix event, our first ever at the historical Orpheum Theatre!
Your VIP ticket includes the following:
Early Entrance
● Beginning at 5 PM, you may enter the main doors on Adams. Doors open at 6:15 PM for all other ticket holders. Pick up your VIP lanyard (required) at the welcome table in the lobby for early entrance.
Meet and Greet
● From 5-5:40 PM, greet this year's entertainers—Carlos Oscar and Adam Trent! Complimentary hors d'oeuvres and bottled non-alcoholic beverages are included. A cash bar, including alcoholic beverages, will be available. A VIP lanyard is required for this event.
The Orpheum Theatre manages the 305 Garage (300 West Washington; $12 flat fee). An alternative is the Wells Fargo Garage at Washington & 1st Avenue.
Thank you for helping us resource the effort against commercial sexual exploitation through your VIP ticket purchase. I am looking forward to an amazing night of world-class entertainment for justice!
Brad Pellish


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Originally posted by ollncasino
reply to post by aaron2209
Many people assume that Islam is a religion of peace corrupted by a misguided minority.
In fact Muhammad authorized the assassination of people who criticized Islam, according to the first biography on his life written by a devout Muslim in 768 AD.
• Muhammad authorised the murder of a number of poets (male & female) who had mocked him. One poet was a mother sleeping with her young children. The Muslim assassin had to move the child aside before thrusting his dagger into her chest. Another victim was a male poet who was, according to Muslim sources, over 100 years old. Mohammed then absolved the Islamic murderers of any wrong doing.
• He also had a propensity to execute prisoners who had been political opponents in the past. One such man had fallen foul of Muhammad when Muhammad was in his early days due to being a better story teller than Muhammad,. "Who will look after my young daughter?" cried the man, moments before he was executed. Muhammad then cursed him.
The Life of Muhammad (768 AD) by Ibn Ishaq
Sadly, the violent Muslims are the ones emulating its founding prophet in both spirit and actions.
edit on 15-9-2012 by ollncasino because: (no reason given)
Originally posted by aaron2209
Originally posted by ollncasino
many people assume that Islam is a religion of peace corrupted by a misguided minority.
I've never assumed anything about any religion. The tiny bit of knowledge I have about all religions is that they all have their archaic ideologies and mantra and when taken literally are quite barbaric.
I had just hoped, possibly naively, that we were progressing.
Originally posted by aaron2209
How disgusting
I have no other words.....
Originally posted by Nucleardiver
Well if it offends them then don't watch it, its as simple as that.
Originally posted by aaron2209
Originally posted by cartenz
When we deny any group the right to protest; we are denying ourselves accountability to government.
I agree but for me it's that anger towards non-muslims that is the sticking point. As shown above, that sign destroys any legitimacy this protest may have had.
Originally posted by aaron2209
This whole stupid ordeal has played right into the hands of the idiots that made the film.
It's damn sad!
Originally posted by kudegras
We take these people from war torn countries and give them a second chance, and within a few years they start trying to change our views to views of their own.
Now in Sydney they are protesting over an American made film. WTF? To see children carrying banners to behead any that dont agree with their fundamentalist narrow world view makes me sick.
I say send them back, back to their own lands where they dont have to put up with our Western ways they seem so determined to tear down.
Those that want to assimilate and live as Aussies are welcome and those that want to change our ways can take a hike, we dont want and dont need their crap.
Julia take a stand, before one of these morons actually does behead an Australian citizen for holding an opposing view.
Originally posted by munkey66
took a couple of days for the protest to take place, wouldn't have been hard to have a few agent provocateurs make their way in and make the situation a little worse than it should have been.
problem, reaction, solution. and repeat.
Originally posted by detachedindividual
Where does freedom end and incitement begin? If you can argue that the sign being held up is incitement, then isn't the video that started all of this also incitement?
Originally posted by aaron2209
It is testing my tolerance and opinions on Islam when I see signs like "Behead all those who insult the prophet" being carried through Sydney CBD.
Originally posted by munkey66
I mentioned already that being a Muslim is a choice .......
Originally posted by FlyersFan
Originally posted by munkey66
I mentioned already that being a Muslim is a choice .......
Then why is it against the law in Muslim countries to leave Islam?
Why is that Christian pastor on death row because he supposedly 'left Islam'?
Escaping Islam is a dangerous thing for those brave enough to try ....
Between 1860 and the 1890s a number of Central Asians came to Australia to work as "Afghan" camel drivers
The first mosque in Australia was built in 1861 at Marree in South Australia. The Great Mosque of Adelaide was built in 1888 by the descendants of the Afghan cameleers
During the 1870s, Muslim Malay divers were recruited through an agreement with the Dutch to work on Western Australian and Northern Territory pearling grounds. By 1875, there were 1800 Malay divers working in Western Australia. Most returned to their home countries.
1967 and 1971, approximately 10,000 Turks settled in Australia under an agreement between Australia and Turkey.
Originally posted by Nucleardiver
In light of the events of this week it seems that the entire Muslim world has gone even more insane than normal and its all because of a movie that they find "offensive". Well if it offends them then don't watch it, its as simple as that. However these fanatics won't ignore it because they look for any excuse they can find to inflict and spread their hatred and violence.
Originally posted by aaron2209
How disgusting
I have no other words.....
Originally posted by borntowatch
Isnt it funny, everyone bitchin about muslims not respecting Sydney or something.
The movie maker is the cause of the problem, but thats not the issue here.
Blame the muslims thats easiest, its ridiculous.


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Kubiak, Frazier give Rice twin connections to NFL
After Thursday's season opener against visiting UCLA, Corey Frazier and Klein Kubiak will engage in a fun conversation once they reach the Rice locker room.
And it won't be about Rice football. It will be about their dads' fortunes that night.
Leslie Frazier will lead the Minnesota Vikings against Gary Kubiak and the Texans at Reliant Stadium in a preseason finale a half hour before the Owls face the Bruins three miles away.
"My focus is on UCLA, but it's also pretty fun in the locker room getting to chat with Corey," said Klein Kubiak, a junior wide receiver. "We're looking forward to seeing what happens (with the Texans-Vikings game) when we get off this field Thursday."
Rice is the only school that boasts the sons of two current NFL head coaches.
Klein Kubiak said the two do not have a friendly wager riding on Thursday's meeting. He added they may have to come up with something when the teams play during the regular season on Dec. 23 in Houston.
Kubiak, a Strake Jesuit graduate, has caught 23 passes in 23 games in his Rice career. This year, he also will serve as the holder on placements.
Corey Frazier, a native of Eden Prairie, Minn., is a senior safety. Two years ago, he led Rice with 83 tackles. He has made 141 tackles and two interceptions in 35 games.
The players' football relationships with their fathers began differently.
Klein Kubiak said he always welcomed his father as the cornerstone of his football career. He said he and his father talk two or three times a week about football. Whenever he gets a chance, he goes to Reliant to see his dad.
"It's nice to play ball and then come home and have that coach on the side," Klein Kubiak said. "He's watched every game I've played in from little league to now. He's always there for me to tell me what I'm doing wrong and what I'm doing right."
Even though Leslie Frazier was the starting cornerback for the Chicago Bears when they won Super Bowl XX, Corey Frazier said he did not want to listen to his father when he was younger. Corey eventually figured it out and now talks to his dad as often as possible.
"I'm always in his ear and trying to watch film with him," Corey Frazier said. "He doesn't have to (be the first to) tell me what I'm doing wrong. I'm always asking him first what I'm doing wrong."
Gary Kubiak said he and Leslie Frazier have a relationship because of their sons' both playing at Rice. On Thursday that takes a back seat, but only because the timing is bad.
"I'm sure we won't get to see much of it," Gary Kubiak said. "It is interesting. I'm sure Leslie (Frazier) would love to see his boy. He's a heck of a player over there, but we've got work to do - put our team together here and finish up the preseason."
Corey Roepken is a freelance writer.


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Should there be an Individual Flat Tax? The Pro Side Introduction While there are a number of different ways to debate the appropriateness of an income tax system, most academicians would agree that there are four broad categories that encompass the basic principles of a good tax system – fairness, ease, minimal interference and conduciveness to economic growth, and adequacy. Is the tax fair? Fairness at the basic level suggests that all people who are in similar circumstances should pay similar taxes. Under this principle of 'equal treatment of equals', one would expect families with the same ability to pay to have the same tax liability. Though not everyone will agree on the definition of fairness, most Americans will accept as fair the principle that the poor pay nothing, the middle class something, and the rich the highest percentage. Is the tax easy to comply with and to administer? Whether a tax system is easy to comply with and easy to administer is a matter of subjectivity. In judging a system against this principle, however, one should examine the costs of compliance by taxpayers (out of pocket expenses such as tax preparation fees and legal fees as well as time spent on compliance) and the government's costs of administering the system. Complexity in a tax system has a tendency to feed upon itself, making efforts to produce a simpler and more equitable tax system even more complex. First, a progressive personal income tax structure complicates the collection process. Second, many IRS regulations exist solely to combat tax evasion and avoidance. As an example, one need only look at the cost of complying with IRA distribution reporting regulations put in place to assure that distributions are properly reported in income. Third, complexity arises from efforts to influence economic and social behavior. The charitable contribution deduction...
pages: 18 (words: 4927)
comments: 1
added: 10/27/2011
Soviet foreign policy in the 1920's and 1930's was more practical than ideological. Do you agree? In order to properly assess this question we must understand the nature of the Soviet foreign policy within these two decades. When observing the events of this time we can see that a duality of policy existed within the USSR. One was the obvious ideological policy and the other a more discreet practical policy. During the 1920s USSR's foreign policy seemed very ideological, however in reality there were several concealed examples of more practical actions. As time continued throughout the 1930s USSR's foreign policies did not only become more apparent but also more practical and less ideological. The USSR entered the 1920's with a rather traditional policy, stressing the importance of the global revolutionary movement. This was embodied in the Comintern established in 1919. The Comintern or Communist international was introduced with the mission of encouraging global revolutions. Communists worldwide were called to infiltrate armed forces, factories and villages in capitalist countries, to support colonial revolts, and to mainly support Soviet Russia. As a foreign policy this affected many Western European democratic countries that feared that the Comintern might actually succeed in starting a revolution and therefore disrupt the balance of power. Another example of an ideological policy was USSR's relationship to China. A small Chinese Communist party was formed in Shanghai. Even though the Comintern had ideological aims it used a practical method to achieve them. This was done by advising the Communist party to ally with the Chinese Nationalist party. This peculiar allegiance was needed to fight against foreign influence and armed warlords who ruled China at the time. This policy turned out to be a catastrophe. After several successes in Northern China, the Nationalists turned on their allies killing thousands of Communists in...
pages: 5 (words: 1206)
comments: 1
added: 11/02/2011
In what ways is the focus of Changing Perspective demonstrated in the chosen texts? The texts I chose for this essay were: 1. The Road Not Taken by Robert Frost 2. Out, Out- by Robert Frost 3. The Door from stimulus booklet 4. Sky-High from stimulus booklet 5. What Its Like – by Everlast (song) 6. American Beauty the film In all of the chosen texts, the focus of changing perspective is demonstrated. Through use of figurative language, symbols, tone, imagery and key themes, their composers have created pieces of literature that show many facets of life in more than one perspective. From "The Road Not Taken" by Robert Frost, one of the most renowned poets of our time to "What Its Like" by Everlast, a contemporary rock music artist, all of the texts I have chosen represent the focus of changing perspective. In "The Road Not Taken" Frost depicts a scene somewhere in the persona's past. The persona is standing at a fork in the path he is walking down and must make a choice of either one or the other. The persona knows that he will never be able to go back and make the choice again and therefore he chose the path that was "less travelled by". This poem can be interpreted in many ways and that is where the focus of changing in perspective comes in. Firstly, we can take this poem as a symbolism of the greatest journey of all, life. The persona then is Frost himself and he is recounting his life and how he made a decision to go down a road that was less travelled by and that has made all the difference in his life. Also, this poem acts as a guide for all people who are making choices in life as Frost tells of how "way leads onto way" and...
pages: 11 (words: 2821)
comments: 1
added: 10/27/2011
Queen Maatkare Hatshepsut, Pharaoh of Egypt during the 18th dynasty, from 1473 BC to 1458 BC, was one of only a handful of female rulers of ancient Egypt. Her story is unique in Egyptian history, and has been the source of many disputes among scholars. Hatshepsut reigned longer than any other female pharaoh. Among the legacies she left behind, none is greater than the mortuary temple she erected at Deir el Bahari in Thebes, the ruins of which still stand in present-day Luxor. The temple, designed by Senenmut, reflects the adjacent mortuary temple of Mentuhotep II, but is much larger. Reliefs and inscriptions on the temple walls tell stories from Hatshepsut's life, and profess her connection to the divine. Based on current knowledge, this essay will provide detailed information about Queen Hatshepsut and her mortuary temple. Hatshepsut was born around 1502 BC to Thutmose I and Ahmose. Both of her parents were from a royal background, and Thutmose I was Pharaoh when she was born. Her two brothers died in accidents, which meant that she was in a position to take over the throne after her father died. This was an unusual situation because very few women had ever become pharaohs. However, Hatshepsut was favored by her parents over her brothers, and she was beautiful and had a charismatic personality. Thus, despite her being a female, she had the makings to become a queen. Thutmose II was Hatshepsut's half-brother and husband, a common situation in ancient Egypt, where brother-sister and father-daughter marriages were accepted. When Thutmose I died, Hatshepsut was about 15 years old, and Thutmose II took over as pharaoh. Thutmose II died after only three or four years of rule, most likely of a skin disease. Hatshepsut had a daughter, named Neferure, but Thutmose II also had a son with...
pages: 7 (words: 1810)
comments: 22
added: 11/02/2011
Discussion On No Real Seperation Of Executive, Legislature, and Judiciary Powers In The UK ConstitutionPolitics
There Is No Real Seperation Of Powers In The UK Constitution Nor Does There Need To Be.'Discuss What are the separated powers? Executive = the administrative branch of government; it makes laws by way of delegated legislation and drafts bills. Legislature = the law making branch of government Judiciary = the law enforcing branch of government What can you say about the separation of powers? The overlap of powers allows Parliament to make any change it wishes by Act of Parliament and helps to insure against arbitrary exercise of power. There is almost complete separation of powers in the USA where governmental intransigence in controversial matters is a much bigger problem than it is here (here the judges can indulge in greater law making activities if necessary). There is little executive-legislature separation in the UK but much legislative-executive/judiciary separation. The concept of the separation of powers was first discussed by Charles Montesquieu (after observing the British system). -------------------------------------------------------------------------------- To what extent do we have imperfect separation of powers? The executive controls, and makes the legislature by creating peers(subject to the approval of the Queen), whips, dissolution (subject to the approval of the Queen, but note that permanent prorogation possible without her permission), prorogation (in theory this is under the Royal Prerogative but the Queen's permission is not necessary), controlling the timetable. Part of the legislature (the government ministers) form the executive. The legislature controls the judiciary by removing senior judges (in the name of the Lord Chancellor). The executive (the Lord Chancellor) controls the judiciary by removing (or perhaps worse not removing) junior judges. The judiciary do not have their salary voted on by Parliament The judiciary reviews the activities of the executive. Paid judges cannot be in the Commons. The executive (the Lord Chancellor (with the Prime Minister in appointing law lords)) appoints judges. The executive makes treaties, which have a degree of legislative influence (though not true legislative...
pages: 4 (words: 1027)
comments: 79
added: 10/27/2011
With any essay that I write, my main objective is to paint a picture with words. There are three basic guidelines that I follow in order to paint this picture. By writing down my ideas, putting my ideas together on a word processor and then going back to revise is how I compose an essay or any other written work. I feel that these guidelines are essential to a well developed essay. When I'm given a writing assignment, one of the first things that I do is focus on the subject or topic of the essay. If it's a subject that I am knowledgeable of, I would add my own input and incorporate it into my writing. And if it's not a subject that I am familiar with, I would do some type of research on the subject. One of the most important processes of my written work is the gathering of detailed ideas. When I am forming ideas about my subject for an essay, the first thing that I usually do is concentrate on what I want to write. What do I want to say to my audience? What is the purpose of my writing? I have learned that by knowing the purpose of your writing keeps you focused on it. Once I have found the answers to these questions, I try to stay focused. I gather details for my subject that will help support and develop it. I try to utilize sensory details, specific examples, facts and statistics, or incidents or anecdotes - if they are needed. The next thing that I do is figure out how do I want to say it. I'll start by writing out a few sentences about my subject and then I determine which one represents my idea most effectively. Once I have found...
pages: 3 (words: 623)
comments: 2
added: 10/08/2011
In Webster's Online Dictionary, the word hero is described as "a man admired for his achievements and noble qualities, one that shows great courage." A modern day hero would be represented through the soldiers who are fighting in the Middle East for America. Those men and women are putting their lives in danger to benefit millions of other people. That is a heroic act that should not go unnoticed. In Maya Angelou's autobiography, I Know Why the Caged Bird Sings, she describes many people whom she felt depicted the word "hero" in all of its glory. I chose to write my essay on three of these main characters in her life: her older brother Bailey Johnson, her mother's boyfriend, Mr. Freeman, and the central character Maya, or Marguerite Johnson. Mr. Freeman is a counter-example of a hero, and the only example that deals with false heroism. Maya never had a father figure in her life, and Mr. Freeman was the only man who ever made her feel wanted. Maya showed her weakness for Mr. Freeman in saying " he held me so softly that I wished he wouldn't ever let me go. I felt so at home. From the way he was holding me I knew he'd never let me go or let anything bad ever happen to me. This was probably my real father and we had found each other at last. (Pg. 73.)" Mr. Freeman gave her a sense of worth, and even though his intentions were cruel and immoral, Maya finally had found what she was longing for. Maya loved Mr. Freeman, but feared him even more. When Mr. Freeman raped Maya when she was eight-years-old, he made sure she would be able to keep the secret from the rest of her family, including her brother Bailey. "If you...
pages: 4 (words: 1008)
comments: 0
added: 01/15/2012
"Karen!", she called, "what am I going to write my essay on!" I looked up to see Abby, my animated friend. I felt the corners of the mouth curl up into the familiar smile as she ran toward me and pressed a leaf into my palm. I looked at it for a moment before giving her a quizzical glance. "I picked it for some inspiration but it isn't helping," she said with a huff and a slight frown. I looked down at the crumpled foliage in my hand and turned it round in my fingers. It's fascinating how every leaf is different. I looked at this one - it was a deep green moving into a hint of yellow along the edges. It was about to change. I thought of me - I was changing too and it was scary to no longer be that little girl I had grown used to but a young lady - eager to experience life and yet nervous of what that meant. Things get harder as one gets older and this leaf looked scared to change too - putting off the inevitable. It had deep wrinkles all along it and as I looked at my hand I saw my wrinkles too. I traced the lines with my gaze as I do sometimes when I look into my grandmothers face. Each choice she had made had created a new line, a new memory, a new lesson. It had a deep cut along one of these lines. I thought how heartbreak could do that to a person - cut so deep that it almost tears them in two. But we are one up on the leaves, I thought triumphantly. We can put the pieces back and stitch them so that they grow back together. I looked at the long...
pages: 3 (words: 574)
comments: 1
added: 10/11/2011
As I began to think about what I wanted to write my paper about, I thought of all the topics in education that I would like to know more about. I finally choose ADD/ADHD and the use of Ritalin as my subject matter. Not being a parent or an experienced educator as of yet, I see America's propensity to over medicate reaching our youngest population. I think the statement by Dr. Breggin is quite profound, "When we drug millions of children to make them more compliant and easier to manage at home and in school, it says more about our society's distorted values than about our children" (Breggin 1998). I know from my experience in the health care profession, that Americans believe in medication to create a better life for themselves. That can be as simple as getting an antibiotic for the slightest cold or as complex as medicating ourselves to fight a chronic condition. We live in a society that wants everything done quickly. We drive through for our food, banking, prescription pick-ups, and car washes. We develop our pictures in an hour and pay for our gas at the pump. We watch 50 channels of television in a second. Television has learned that it has to be gripping in order to keep up with our wandering minds. We even have televisions that allow you to watch more than one channel at a time. We wonder why our children have trouble paying attention. When do we do anything that requires anything but the most miniscule amount of our attention? How do we expect children to practice the learned behaviors of watching and listening, if we never teach it to them? We spend the first few years of their lives showing them the frenzied world in which we live and then...
pages: 15 (words: 4110)
comments: 0
added: 02/19/2012
I chose to write my paper on Jean Piaget's Theory of Cognitive Abilities. I chose this topic because I as well as many people in my family have been labeled "gifted" early on in school. I realize that this is a measure of intelligence defined as the potential for learning, however none of us labeled gifted have done anything more successful than other members of my family. I fact the opposite may be true. Perhaps a gifted labeling made us all lazy. Nonetheless, I would like to know how Piaget's theory relates to how intelligence is measured. Piaget became fascinated early in his studies with the discovery that children of the same age often gave the same incorrect answers to questions, suggesting that there were consistent, qualitative differences in the nature of reasoning of different ages, not simply a quanitive increase in the amount of intelligence or knowledge. This discovery marked the beginning of Piaget's continuing effort to identify changes in the way children think, how they perceive their world in different ways at different points in development. The different stages postulated by Piaget help to explain different rates of learning at different ages as well as the types of learning possible at different ages for the majority of the population. Learning itself is seen by Piaget as a process of discovery on the part of the individual, and learning as a formal activity becomes a system of organization, by which instruction is enhanced by the way the teacher arranges experience. Learning is thus experimental, and Piaget suggests that experiences have meaning to the extent that they can be assimilated. There are two principal learning theories in psychology, one of which focuses on the learning process while the other focuses on ones capacity to learn. Piaget offered a biological theory of intelligence that he presented as a unified approach to intelligence and learning. Piaget restricted the idea of learning to an acquisition...
pages: 3 (words: 676)
comments: 2
added: 12/03/2011


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Quick Reference
In statistics, a non-parametric and distribution-free test of the randomness of an ordered sequence of binary or dichotomous observations, in which the number of runs expected on the basis of chance is calculated and compared with the observed number of runs. Also called the runs test. [Named after the Romanian-born US statistician Abraham Wald (1902–50) and the Polish-born US statistician Jacob Wolfowitz (1910–81) who developed it in 1940]
Subjects: Psychology.
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Remember the original PlayStation 3? Aside from being expensive (the 60GB model went for a heft $599 on launch day), the original model was also ginormous (who can forget all those hilarious PhotoShops that punctuated the system's unveil). And the upcoming PlayStation 4 looks equally monolithic. Or is it?
That's exactly what PlayStation Access decided to find out:
As you can tell, when in a vertical position, the PS3 is actually a tad bit taller than the PS4. But when both are flat, the PS3 ends up being much taller. Otherwise, they both seem equally deep.
Maybe it's just one person's opinion, but the PS4 looks much nicer when in its horizontal position. Whereas vertical, it just seems a tad bit awkward, especially its stand.


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Be the first YellowBot user to add photos or videos of Wandas Beauty Shop
Wanda's Beauty Shop was founded in 1980, and is located at 25771 Hwy 36 in Anton. It employs 1 employees and is generating approximately $20,000.00 in annual revenue.
Wandas Beauty Shop is located at 25771 Us Highway 36, Anton, CO. This business specializes in Beauty Salons.
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Hosted by Mykel Hawke, survival expert and former Green Beret, each episode of One Man Army pit four operators against various military-style challenges to test intelligence, decision-making, endurance, and physical strength.
Devised with the input of active Navy SEALs, this is hands down the most intense challenge I've ever designed. Contestants, armed with only a handsaw, had a small hole at the top of the coffin to take a breath, hold it, then swim to a grate with four inch-thick aluminum bars they needed to saw through to escape. Safety was our number one priority, and a number of failsafes were implemented to evacuate the participant in a matter of seconds.
Four contestants are given 15 seconds to view a maze having one entrance and one exit. They then must crawl through the maze on their back as various distractions occur around them, including explosions, fireballs, water, brush, smoke, sandbags and thorns.
Two contestants are locked in opposing cells with various items such as clothing, hangars, a bed, a chair, etc. The objective is to "MacGuyver" a device that allow them to reach nine feet out of their cell and snag the key to their cell. First one to unlock their cell and strike the bell wins the game.
I had custom ice blocks with items frozen inside made for this speed challenge. Contestants had to obtain 5 keys, but had to decide if it was worth wasting the time breaking the bigger blocks to obtain tools that would make it easier to smash the key blocks.
Custom wall plug with freezer door was built to match the walls of the 40' freezer truck. Contestants needed to break ice blocks to get the 5 keys required to unlock each lock and finally break free of the freezing temperatures inside.
Host Mykel Hawke coaching a contestant through the harrowing obstacles of the maze from his observation post.
Little did we know at the time that this would be the only man out of 8 to successfully escape from the dreaded "Water Coffin."
Originally sketched out on graph paper--with each block representing a foot--made this an easy transition to actual construction. From this vantage point, can you find the exit?


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DENVER, June 21, 2001 — Increased levels of the stress hormone cortisol are clearly elevated in the bloodstream of child-bearing-aged women who have stopped menstruating, a senior researcher at the Magee-Womens Research Institute has found. The study is significant because it also shows that a psychological intervention and stress-reduction therapy can have a positive impact on infertility associated with the cessation of menstruation.
Called functional hypothalamic amenorrhea (FHA), the condition affects some 5 percent of women in their reproductive years. It is characterized by wildly irregular or absent periods – often for as long as two years or more.
"FHA isn't caused by any real defects in the reproductive system," said Sarah L. Berga, M.D., a professor in the departments of obstetrics, gynecology and reproductive sciences and psychiatry at the University of Pittsburgh School of Medicine. "We thought using stress-reduction techniques might reverse the process, and our results show we were right."
Dr. Berga presented the study findings at the 83rd Annual Meeting of the Endocrine Society, an organization made up of endocrine specialists in internal medicine, pediatrics and gynecology.
Now in its seventh year, the ongoing study has included both human volunteers and primates to link stress and infertility due to FHA. "We are finding out how stress causes infertility," said Dr. Berga, who is also director of the division of reproductive endocrinology and infertility at the Magee-Womens Hospital of the University of Pittsburgh Medical Center.
"This is something I've been working on for 15 years," she said. "Typically, these women are not aware they're under stress, or don't acknowledge it."
Women with FHA also share certain characteristics, such as a tendency toward perfectionism and poor nutritional or lifestyle choices. Many just try to get too much done in a day.
Fourteen women took part in the most recent phase of the study. Seven were treated with sessions of cognitive behavior therapy and dietary counseling, while seven were not. All were observed for a period of 20 weeks.
Of the treated women, six fully recovered normal menstrual cycles, and one partially recovered. Of those not treated, only one fully recovered. One partially recovered, and five continued to experience FHA.
"The endocrine system rewards moderation," explained Dr. Berga. "We try to teach balance and creative problem solving."
Long-term FHA affects more than fertility. It also can result in bone loss and affect brain function. If women with FHA take fertility drugs to get pregnant, the underlying stress process may lead to early delivery or poor fetal brain development.
"Without intervention, these women are at risk for a recurrence," Dr. Berga said. "But the good news is that treatment is effective."
In addition to Dr. Berga, study authors include Tammy L. Loucks, M.P.H.; Marijane Krohn, Ph.D.; Judy L. Cameron, Ph.D; and Marsha D. Marcus, Ph.D.
Funding for the study was provided by the National Institute of Mental Health, a component of the National Institutes of Health.
Magee-Womens Research Institute is the country's first research institute devoted to women and infants. It was formed in 1992 by Magee-Womens Hospital of University of Pittsburgh Medical Center. The University of Pittsburgh School of Medicine's department of obstetrics, gynecology and reproductive sciences is one of the top three departments in the nation in NIH funding.
For more information on FHA or this research study, please call 412-641-1602.
Affiliated with the University of Pittsburgh Schools of the Health Sciences
Supplemental content provided by Healthwise, Incorporated. To learn more, visit www.healthwise.org
For help in finding a doctor or health service that suits your needs, call the UPMC Referral Service at 412-647-UPMC (8762) or 1-800-533-UPMC (8762). Select option 1.
UPMC is an equal opportunity employer. UPMC policy prohibits discrimination or harassment on the basis of race, color, religion, ancestry, national origin, age, sex, genetics, sexual orientation, gender identity, marital status, familial status, disability, veteran status, or any other legally protected group status. Further, UPMC will continue to support and promote equal employment opportunity, human dignity, and racial, ethnic, and cultural diversity. This policy applies to admissions, employment, and access to and treatment in UPMC programs and activities. This commitment is made by UPMC in accordance with federal, state, and/or local laws and regulations.
Medical information made available on UPMC.com is not intended to be used as a substitute for professional medical advice, diagnosis, or treatment. You should not rely entirely on this information for your health care needs. Ask your own doctor or health care provider any specific medical questions that you have. Further, UPMC.com is not a tool to be used in the case of an emergency. If an emergency arises, you should seek appropriate emergency medical services.
Pittsburgh, PA, USA | UPMC.com


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Thieves grab garden gear as nights get lighter
British Summertime sees a 20% rise in shed and garages thefts
With the long-awaited British Summertime just around the corner, homeowners are being warned to lock their garden sheds and garages as light-fingered thieves will be out to take advantage of the extra daylight.
Analysis of ten years of our claims data* reveals that thefts from gardens and outbuildings shoot up by 20% when the clocks go forward with the average haul totalling around £850.
And thieves will be keen to get their hands on anything within easy reach – from children's toys to bikes to garden tools:
Of the garden equipment stolen last year:
- 42% of thefts were for power tools, of which 56% were drills
- 39% were for simple hand tools like spades, shovels, rakes and forks
- 19% were for electric and petrol lawnmowers, chainsaws and ladders**
However, the thieves don't just wait for British Summertime to begin before they start looting gardens, as the lighter evenings in March show a 10% increase in thefts compared with the dark nights of January.
And as the nights get lighter the number of claims increase, with July and August peeking at almost 30% higher than at the start of the year.
Jonathan Cracknell, our household underwriting manager, says; "We're all looking forward to a bit more sunshine, but lighter evenings also mean all your expensive power tools, bikes and children's toys are more visible to passing criminals.
"So after a day in the garden or doing a bit of DIY ensure everything is secured away inside your home, shed or garage. And remember items such as ladders and wheelie bins are perfect for burglars to use to get into your home, so keep them out of reach and out of sight if you can."
Our cover
Our household policies provide cover for theft of contents in locked/secured outbuildings, garages and garden sheds up to £2500, so that would include your mountain bike, lawnmower and set of golf clubs.
Up to £500 of your contents in the garden are also covered, as standard, for theft and damage caused by things like fire, malicious damage or extreme weather events. This includes patio furniture, pots and BBQs, but if you have some particularly nice garden ornaments or a fancy table and chairs that cover can be increased to £1500 from around £9 a year.
But remember bikes are not covered if you leave them lying in your garden.
If you do want cover for your bikes outside the home and anywhere in the world you might want to take out extra cycle cover which again costs from around £9 a year for the whole family's bikes. (Cover limits from £300 to £2,500)
Similarly if you would like your golf clubs or fishing tackle to be covered outside the home you could consider taking out additional personal belongings cover, which again covers you anywhere in the world. It costs from around £27 a year for £2000 worth of cover.
Our Top Tips to beat the shed burglar
- Don't tempt the thieves in, put away your garden tools, paddling pool, trampoline and bikes at the end of the day
- Lock your sheds and out-buildings, replace any rusty padlocks.
- Close gates and repair damaged fences - deterrents are sometimes the best prevention.
- Hide or cover bulky items which can't be locked away, such as large gas barbecues and picnic benches.
- Lock up ladders and secure wheelie bins – they are perfect for giving burglars a leg up into the rest of your home!
* Aviva claims data 2003 – 2012
** Aviva theft claims data from January to December 2012


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Joe McKnight, star tailback of USC's football team, has been driving a sport utility vehicle owned by a Santa Monica businessman, an arrangement the school is investigating and which may be in violation of NCAA rules.
The NCAA prohibits student-athletes from accepting benefits from marketing representatives or agents or "extra benefits" based on their athletic ability.
For several weeks, McKnight has been seen driving a well-kept 2006 Land Rover that, according to California Department of Motor Vehicles records, is registered to Scott Schenter.
Schenter, 47, has a background that includes marketing, and a company he owned had previously registered a website called 4joemcknight.com. It is unclear whether Schenter has any ties with USC or whether the school had knowledge of his activities.
If it's found that McKnight is in violation of NCAA rules, it could affect his athletic eligibility. USC's football team will next play against Boston College in the Emerald Bowl on Dec. 26 in San Francisco.