Significant insight has emerged from the analyses of phylogenetics, compensatory base-pair mutations, and photochemical crosslinking that indicate a specific catalytic role for U6 small nuclear RNA (snRNA) in pre-messenger RNA splicing. This notion has been strengthened by our recent discovery that the 3' intramolecular stem-loop of U6 snRNA binds a magnesium ion that is critical for the chemistry of splicing (Yean et al., Nature 408:881). It is unclear, however, whether the magnesium that we have identified is at the catalytic site and how the 3' stem-loop of U6 interacts with the rest of the spliceosome. The objectives of this pre-doctoral grant application arc two-fold: (1) test a metal-ligand interaction that would involve juxtaposing the 3' intramolecular stem loop (ISL) of U6 snRNA with the 5' splice site (SS) and (2) to investigate what protein factors stabilize, assist, or mediate RNA folding that leads to catalysis within the context of the assembled spliceosome. These interactions, once defined, would help our understanding of how the different sites of chemistry become tightly linked during spliceosome catalysis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31GM067579-03
Application #
6784560
Study Section
Special Emphasis Panel (ZRG1-F05 (29))
Program Officer
Toliver, Adolphus
Project Start
2002-09-30
Project End
2005-09-29
Budget Start
2004-09-30
Budget End
2005-09-29
Support Year
3
Fiscal Year
2004
Total Cost
$26,847
Indirect Cost
Name
City of Hope/Beckman Research Institute
Department
Type
DUNS #
027176833
City
Duarte
State
CA
Country
United States
Zip Code
91010
Dery, Kenneth J; Yean, Shyue-Lee; Lin, Ren-Jang (2008) Assembly and glycerol gradient isolation of yeast spliceosomes containing transcribed or synthetic U6 snRNA. Methods Mol Biol 488:41-63