Abstract

The goal of this research is to extend our knowledge of tRNA gene organization and expression in Escherichia coli. Specific tRNA genes which ave been isolated using recombinant DNA methodologies will be studied with regard to primary structure, then utilized for detailed studies on gene expression in vivo and in vitro. Specific tRNA leu regulatory sequences will be utilized in vitro transcription systems in order to determine what factors regulate tRNA gene expression. These controlling elements will be fused in vitro to the galactokinase gene of E. coli and utilized for detailed in vivo and in vitro studies thus simplifying physiological and genetic analysis. In vitro mutagenesis of these sequences will be employed to critically assess promoter and terminator structure-function relationships. This work impinges on the following major unanswered questions: How are all the tRNA genes of a cell coordinately expressed to ensure faithful protein synthesis? What are the general structural features of tRNA genes and their regulatory sequences? What factors are involved in the controlled initiation and termination of tRNA gene transcription? Ultimately these studies should permit the engineering of modified tRNA genes which will be useful in understanding how tRNA functions in the cell.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025871-07
Application #
3273393
Study Section
(MG)
Project Start
1978-12-01
Project End
1986-11-30
Budget Start
1984-12-01
Budget End
1985-11-30
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Virginia Commonwealth University
Department
Type
Overall Medical
DUNS #
City
Richmond
State
VA
Country
United States
Zip Code
23298

  Publications

Bauer, B F; Elford, R M; Holmes, W M (1993) Mutagenesis and functional analysis of the Escherichia coli tRNA(1Leu) promoter. Mol Microbiol 7:265-73
Wahab, S Z; Rowley, K O; Holmes, W M (1993) Effects of tRNA(1Leu) overproduction in Escherichia coli. Mol Microbiol 7:253-63
Rowley, K B; Elford, R M; Roberts, I et al. (1993) In vivo regulatory responses of four Escherichia coli operons which encode leucyl-tRNAs. J Bacteriol 175:1309-15
Holmes, W M; Andraos-Selim, C; Roberts, I et al. (1992) Structural requirements for tRNA methylation. Action of Escherichia coli tRNA(guanosine-1)methyltransferase on tRNA(1Leu) structural variants. J Biol Chem 267:13440-5
Vnencak-Jones, C L; Wahab, S Z; Zehner, Z E et al. (1987) A human tRNA(iMet) gene produces multiple transcripts. Mol Cell Biol 7:4134-8