Our goals are to understand ribosome function and structure at the molecular level. We plan to use the recently developed method of DNA hybridization electron microscopy to map the locations of specific regions of rRNA, as well as immunoelectron microscopy to map the locations of ribosomal proteins and the spatial arrangement of functional sites on the ribosome. By relating these sites to our knowledge of ribosome structure, and to the ribosomes of diverse organisms, we hope to gain a detailed understanding of the mechanism of protein synthesis. Our objective will be to relate the three dimensional structure of the ribosome to its function during protein synthesis. The small subunit is responsible for recognizing the initiation site on mRNA with the participation of initiation factors and fmet- tRNA, for binding aminoacyl tRNAs, for associating with the large subunit, and for regulating the translational fidelity of messenger reading. The large subunit binds the acceptor stem of aminoacyl tRNAs entering the A site; catalyzes peptidyl transfer, and participates in elongation and translocation. By relating the three dimensional distributions of ribosomal proteins, factors, and regions of RNA's with known biochemical information, we will attempt to elucidate the structural aspects of the molecular events occurring during protein synthesis. Comparative studies of ribosome structure will also be pursued in order to relate structural features of prokaryotic, eukaryotic, and organellar ribosomes to their common functions in protein synthesis. Because these studies are so broad and relate to fundamental cellular mechanisms, we expect that they will be basic to all aspects of human health. Protein synthesis is a central part of the mechanism of all cells, and we hope our results can be broadly useful in diverse endeavors that range from treating diseased or abnormally growing human cells to controlling bacterial infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM024034-13
Application #
3272032
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1978-12-01
Project End
1992-06-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
13
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Type
Schools of Arts and Sciences
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Tsay, Y F; Shankweiler, G; Lake, J et al. (1994) Localization of Saccharomyces cerevisiae ribosomal protein L16 on the surface of 60 S ribosomal subunits by immunoelectron microscopy. J Biol Chem 269:7579-86
Oakes, M I; Lake, J A (1990) DNA-hybridization electron microscopy. Localization of five regions of 16 S rRNA on the surface of 30 S ribosomal subunits. J Mol Biol 211:897-906
Oakes, M I; Kahan, L; Lake, J A (1990) DNA-hybridization electron microscopy tertiary structure of 16 S rRNA. J Mol Biol 211:907-18
Oakes, M; Scheinman, A; Rivera, M et al. (1987) Evolving ribosome structure and function: rRNA and the translation mechanism. Cold Spring Harb Symp Quant Biol 52:675-85
Lake, J A (1987) Prokaryotes and archaebacteria are not monophyletic: rate invariant analysis of rRNA genes indicates that eukaryotes and eocytes form a monophyletic taxon. Cold Spring Harb Symp Quant Biol 52:839-46
Lake, J A (1987) Determining evolutionary distances from highly diverged nucleic acid sequences: operator metrics. J Mol Evol 26:59-73
Lake, J A (1987) A rate-independent technique for analysis of nucleic acid sequences: evolutionary parsimony. Mol Biol Evol 4:167-91
Reisler, E; Cheung, P; Borochov, N et al. (1986) Monomers, dimers, and minifilaments of vertebrate skeletal myosin in the presence of sodium pyrophosphate. Biochemistry 25:326-32
Langer, J A; Lake, J A (1986) Elongation factor Tu localized on the exterior surface of the small ribosomal subunit. J Mol Biol 187:617-21
Oakes, M I; Clark, M W; Henderson, E et al. (1986) DNA hybridization electron microscopy: ribosomal RNA nucleotides 1392-1407 are exposed in the cleft of the small subunit. Proc Natl Acad Sci U S A 83:275-9

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