Transfer RNAs are essential components of all biological systems. Although the role of tRNA in protein synthesis is usually emphasized it is important to recognize that the molecule is involved in a variety of other biological functions: reverse transcription of RNA to DNA in the retroviruses; regulation of enzyme synthesis; regulation of cell division; and changes in tRNA populations during embryogenesis and oncogenesis. It is not clear why the tRNA molecule is used in such a large variety of biological functions or how the sequence diversities necessary to achieve these functions are generated. This proposal implements genetics, biochemistry, and statistics as converging operations to analyze tRNA, therein allowing the disclosure of unprecedented information about the molecule. We propose to elucidate: - the nucleotide domains of the 20 tRNA classes responsible for their amino acid specificities; - the merits of our scheme for the origin and evolution of primitive tRNA molecules (continuing 9-19 nucleotides) that encoded 4 classes of amino acids distinguished only grossly by the general chemical nature of their functional side chains. By extention, the statistical and mathematical methods we develop can readily be adapted to a number of important questions related to health problems. For example, they are readily adaptable to influenza viruses to determine how the 8 genomic RNAs necessary for flus' viral activity are recognized as distinct during virion assembly. Such knowledge could then be used to prevent assembly (and thus disease) in a manner that does not adversely impact on the physiology of the host cell(s). Also amenable to study (with potential for cure) is the assembly of the oncogenic retrovirus, for their virions contain one specific tRNA and two """"""""35S genomic"""""""" RNAs.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM042123-18
Application #
3300667
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1988-08-01
Project End
1993-07-31
Budget Start
1990-08-01
Budget End
1991-07-31
Support Year
18
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Earth Sciences/Natur
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
McClain, William H (2006) Surprising contribution to aminoacylation and translation of non-Watson-Crick pairs in tRNA. Proc Natl Acad Sci U S A 103:4570-5
Lee, Dennis; McClain, William H (2004) Aptamer redesigned tRNA is nonfunctional and degraded in cells. RNA 10:7-11
McClain, William H; Gabriel, Kay; Lee, Dennis et al. (2004) Structure-function analysis of tRNA(Gln) in an Escherichia coli knockout strain. RNA 10:795-804
Choi, Hyunsic; Gabriel, Kay; Schneider, Jay et al. (2003) Recognition of acceptor-stem structure of tRNA(Asp) by Escherichia coli aspartyl-tRNA synthetase. RNA 9:386-93
Choi, H; Otten, S; McClain, W H (2002) Isolation of novel tRNA(Ala) mutants by library selection in a tRNA(Ala) knockout strain. Biochimie 84:705-11
Choi, Hyunsic; Otten, Sharee; Schneider, Jay et al. (2002) Genetic perturbations of RNA reveal structure-based recognition in protein-RNA interaction. J Mol Biol 324:573-6
McClain, W H; Gabriel, K (2001) Construction of an Escherichia coli knockout strain for functional analysis of tRNA(Asp). J Mol Biol 310:537-42
Gabriel, K; McClain, W H (2001) Plasmid systems to study RNA function in Escherichia coli. J Mol Biol 310:543-8
Moulinier, L; Eiler, S; Eriani, G et al. (2001) The structure of an AspRS-tRNA(Asp) complex reveals a tRNA-dependent control mechanism. EMBO J 20:5290-301
Biala, E; McClain, W; Strazewski, P (1999) Thermodynamics of site-specific variant tRNA(Ala) acceptor stem microhairpins. Nucleosides Nucleotides 18:1575-6

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