Knowledge about the three dimensional structure of ribonucleic acids (RNAs) and ribonucleoprotein particles (RNPs) is essential for understanding how they carry out their biological functions, which include RNA processing and the translation of the genetic information of messenger RNA (mRNA) into protein sequences. Such understanding could offer opportunities for pharmacological or genetic intervention in these critical processes. This proposal describes the development of three dimensional models for RNAs and RNPs, with emphasis on the ribosome and tmRNA. The ribosome is the site of protein synthesis in the cell, while tmRNA is the product of the ssrA gene in bacteria. tmRNA combines functions of both transfer RNA (tRNA) and mRNA and is used to terminate translation of messages lacking a stop codon, to release the ribosome for recycling, and to tag the truncated protein product for degradation by cellular proteases. The studies on ribosomal structure/function relationships will incorporate all available high resolution data from x-ray crystallography and nuclear magnetic resonance into models matching the density maps from cryo-electron microscopic (cryo-EM) reconstructions of ribosomes trapped at various points in the translational cycle. Dr. Harvey seeks to understand the structural, energetic and kinetic basis of translational fidelity and of translocation. The tmRNA research will include studies of the molecule free in solution, using transient electric birefringence and molecular modeling, along with cryo-EM investigations of tmRNA structure / function relationships on the ribosome. This research will include development of a library of RNA structural motifs derived from the Nucleic Acid Database, along with tools for incorporating these motifs into structural models.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM053827-06
Application #
6328411
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Program Officer
Lewis, Catherine D
Project Start
1995-09-30
Project End
2005-03-31
Budget Start
2001-04-01
Budget End
2002-03-31
Support Year
6
Fiscal Year
2001
Total Cost
$204,375
Indirect Cost
Name
University of Alabama Birmingham
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294
Taylor, Derek J; Devkota, Batsal; Huang, Andrew D et al. (2009) Comprehensive molecular structure of the eukaryotic ribosome. Structure 17:1591-1604
Mears, Jason A; Sharma, Manjuli R; Gutell, Robin R et al. (2006) A structural model for the large subunit of the mammalian mitochondrial ribosome. J Mol Biol 358:193-212
Stagg, Scott M; Harvey, Stephen C (2005) Exploring the flexibility of ribosome recycling factor using molecular dynamics. Biophys J 89:2659-66
Stagg, Scott M; Mears, Jason A; Harvey, Stephen C (2003) A structural model for the assembly of the 30S subunit of the ribosome. J Mol Biol 328:49-61
Gao, Haixiao; Sengupta, Jayati; Valle, Mikel et al. (2003) Study of the structural dynamics of the E coli 70S ribosome using real-space refinement. Cell 113:789-801
Mears, Jason A; Cannone, Jamie J; Stagg, Scott M et al. (2002) Modeling a minimal ribosome based on comparative sequence analysis. J Mol Biol 321:215-34
Stagg, Scott M; Valle, Mikel; Agrawal, Rajendra K et al. (2002) Problems with the transorientation hypothesis. RNA 8:1093-4
Stagg, S M; Frazer-Abel, A A; Hagerman, P J et al. (2001) Structural studies of the tRNA domain of tmRNA. J Mol Biol 309:727-35
VanLoock, M S; Agrawal, R K; Gabashvili, I S et al. (2000) Movement of the decoding region of the 16 S ribosomal RNA accompanies tRNA translocation. J Mol Biol 304:507-15
Wang, R; Alexander, R W; VanLoock, M et al. (1999) Three-dimensional placement of the conserved 530 loop of 16 S rRNA and of its neighboring components in the 30 S subunit. J Mol Biol 286:521-40

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