Regulation of tRNA Gene Expression
{'Code': '08070400', 'Directorate': {'Abbreviation': 'BIO', 'LongName': 'Direct For Biological Sciences'}, 'Division': {'Abbreviation': 'MCB', 'LongName': 'Div Of Molecular and Cellular Bioscience'}}
{'SignBlockName': 'Marcia Steinberg', 'PO_EMAI': None, 'PO_PHON': None}
The relative abundance of different tRNAs is statically                         regulated by the sequence and spacing of internal control                       regions (ICR), both 5' and 3' - flanking sequences, and the                     copy number of individual tRNA genes.  A protein factor,                        TFIIIC, nucleates formation of transcription complex.                           Variations from canonical sequences and spacing in the ICR                      are known to affect this interaction which may determine                        which genes are expressed.  We have participated in                             development of a photosensitive cross-linking reagent, 5-N3-                    deoxyuridine-5'-triphosphate (AdUTP), which can be                              enzymatically incorporated into DNA in place of thymidine.                      The photoactive DNA is capable of sequence-specific                             interaction with TFIIIC and can be used to identify the                         subunit of this protein which contacts the B-block of the                       ICR.  We propose to study the interactions of this protein                      with tRNA genes to identify which T residues are in close                       association with TRIIIC,  whether A &amp; B blocks contact the                      same or different subunits, whether other T residues in the                     coding or flanking sequences can be cross-linked to the                         factor, and what regions of the protein contact the ICR.  The                   gene(s) for the DNA binding subunit (s) will be identified in                   a yeast genomic library using synthetic DNA probes based on                     partial amino acid sequence analysis of purified                                polypeptides.  The nucleotide sequence of this gene (s) will                    be determined to deduce the amino acid sequence analysis of                     purified polypeptides.  The nucleotide sequence of this gene                    (s) will be determined to deduce the amino acid sequence.                       Peptide mapping of TFFIIIC photocrosslinked to AdUTP                            substituted DNA will allow us to identify sites in the                          protein which are in close contact with each tRNA control                       region.  Studies of  mutations in 5'-flanking and anticodon                     sequences which affect tRNA gene expression will also be                        completed.                                                                                                                                                      These investigations deal with the factors that control the                     formation of transfer RNA's, (tRNA's), which are ribonucleic                    acid molecules involved in transferring amino acids to                          nascent peptide chains.  One of these factors is a protein                      called TFIIIC that reacts with the genes that code for these                    tRNA's.  The investigations will employ a base that will                        substitute for thymine in DNA and react with TFIIIC in the                      presence of light.  In this way, the site on the DNA with                       which TFIIIC reacts can be identified.
{'FirstName': 'Jerry', 'LastName': 'Johnson', 'PI_MID_INIT': 'D', 'PI_SUFX_NAME': None, 'PI_FULL_NAME': 'Jerry D Johnson', 'EmailAddress': 'jdjohn@uwyo.edu', 'NSF_ID': '000340694', 'StartDate': '10/15/1988', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'University of Wyoming', 'CityName': 'LARAMIE', 'ZipCode': '820712000', 'PhoneNumber': '3077665320', 'StreetAddress': '1000 E UNIVERSITY AVE', 'StreetAddress2': None, 'CountryName': 'United States', 'StateName': 'Wyoming', 'StateCode': 'WY', 'CONGRESSDISTRICT': '00', 'CONGRESS_DISTRICT_ORG': 'WY00', 'ORG_UEI_NUM': 'FDR5YF2K32X5', 'ORG_LGL_BUS_NAME': 'UNIVERSITY OF WYOMING', 'ORG_PRNT_UEI_NUM': 'FDR5YF2K32X5'}
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[{'Code': '1144', 'Text': 'Molecular Biophysics'}, {'Code': '9120', 'Text': 'CROSS-DIRECTORATE PROGRAMS'}]
{'url': 'https://www.nsf.gov/awardsearch/download?DownloadFileName=1988&All=true', 'xml': '8802058.xml'}