Small ribonucleoprotein particles (snoRNPs) are protein-RNA assemblies that catalyze specific chemical modifications on ribosomal RNA, small nuclear RNA, transfer RNA and some messenger RNA. A common modification required for ribosomal RNA biogenesis is 2'-O-methylation that is catalyzed by box C/D snoRNPs. Currently very little is known about the molecular mechanism of assembly and catalysis of box C/D snoRNPs. A multidisciplinary approach will be undertaken to study the assembly and catalytic function of box C/D snoRNPs that employs crystallographic, biochemical, and mass spectroscopic methods. The following specific aims will be achieved during the proposed period: 1) understand protein-protein interactions required for snoRNP assembly both by determining a crystal structure of the protein complex of two core snoRNP proteins and by the hydrogen/deuterium exchange mass spectroscopy; 2) identify and characterize the catalytic subunit(s) of the box C/D snoRNP by crystallographic as well as biophysical binding studies on binding of the predicted methyl donor, S-adenosyI-L-methionine; 3) understand the interaction between the snoRNP proteins and the box C/D RNA required for assembly by biophysical binding studies, co-crystal structure determination, as well as mutagenesis studies between each protein and the box C/D RNA; and 4) obtain a crystal structure of the intact snoRNP including a methylation RNA target. This study will make significant contributions to our understanding of ribosome biogenesis and gene regulation. It also has direct implications in understanding the molecular mechanisms for several diseases linked to the function of box C/D snoRNPs.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM066958-02
Application #
6699993
Study Section
Biophysical Chemistry Study Section (BBCB)
Program Officer
Lewis, Catherine D
Project Start
2003-07-01
Project End
2008-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
2
Fiscal Year
2004
Total Cost
$240,915
Indirect Cost
Name
Florida State University
Department
Physiology
Type
Schools of Arts and Sciences
DUNS #
790877419
City
Tallahassee
State
FL
Country
United States
Zip Code
32306
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