The goals are to elucidate the structures, genomic arrangement, and expression of human transfer RNA genes. Clones of these genes will be isolated by recombinant DNA techniques using 32P-labeled mammalian tRNAs as hybridization probes to screen human gene libraries. The tRNA genes will be characterized by restriction mapping and Southern hybridization techniques, genomic and chromosome-specific blotting experiments, DNA sequence analyses, and transcription studies.
Specific aims for this project are: to elucidate the nature of internal promoter and 5'-flanking regulatory elements for the initiation of transcription of human tRNA genes by RNA polymerase III; to isolate human tRNA genes which have intervening sequences and study the post-transcriptional splicing of precursor tRNAs to the mature-sized tRNAs; to study the biosynthesis of human tRNAs by RNA sequence analyses of the various tRNA intermediates generated using human cell-free extracts; and to construct human mutant tRNA genes by in vitro mutagenesis for transcription studies, for the accumulation of altered precursor tRNAs for processing and modification studies, and for studies designed to elucidate the relationships of tRNA primary structure to specific biological functions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM030671-04
Application #
3278483
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1983-03-01
Project End
1989-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of North Texas
Department
Type
Schools of Arts and Sciences
DUNS #
City
Denton
State
TX
Country
United States
Zip Code
76203
Craig, L C; Pirtle, I L; Gracy, R W et al. (1991) Characterization of the transcription unit and two processed pseudogenes of chimpanzee triosephosphate isomerase (TPI). Gene 99:217-27
Mitchell, A; Bale, A E; Wang-Ge, M et al. (1991) Localization of a DNA segment encompassing four tRNA genes to human chromosome 14q11 and its use as an anchor locus for linkage analysis. Genomics 11:1063-70
McBride, O W; Pirtle, I L; Pirtle, R M (1989) Localization of three DNA segments encompassing tRNA genes to human chromosomes 1, 5, and 16: proposed mechanism and significance of tRNA gene dispersion. Genomics 5:561-73
Shortridge, R D; Johnson, G D; Craig, L C et al. (1989) A human tRNA gene heterocluster encoding threonine, proline and valine tRNAs. Gene 79:309-24
Craig, L C; Wang, L P; Lee, M M et al. (1989) A human tRNA gene cluster encoding the major and minor valine tRNAs and a lysine tRNA. DNA 8:457-71
Pirtle, I L; Shortridge, R D; Pirtle, R M (1986) Nucleotide sequence and transcription of a human glycine tRNAGCC gene and nearby pseudogene. Gene 43:155-67
Shortridge, R D; Pirtle, I L; Pirtle, R M (1986) IBM microcomputer programs that analyze DNA sequences for tRNA genes. Comput Appl Biosci 2:13-7
Chang, Y N; Pirtle, I L; Pirtle, R M (1986) Nucleotide sequence and transcription of a human tRNA gene cluster with four genes. Gene 48:165-74
Johnson, G D; Pirtle, I L; Pirtle, R M (1985) The nucleotide sequence of tyrosine tRNAQ* psi A from bovine liver. Arch Biochem Biophys 236:448-53
Shortridge, R D; Pirtle, I L; Pirtle, R M (1985) Nucleotide sequence and transcription of a gene encoding human tRNAGlyCCC. Gene 33:269-77