The broad aims of our research are to understand the molecular mechanisms and physiological consequences of regulating eukaryotic gene expression by post-transcriptional processes. A large amount of important biological regulation occurs post-transcriptionally, but many of the mechanisms are not well understood. We will study selective mRNA degradation, the processes by which specific mRNAs are actively eliminated from cells. Regulated turnover of mRNAs profoundly influences gene expression, and our broad goals are to understand the mechanisms and consequences of that regulation. We focus on the accelerated degradation of mRNAs that contain premature translation termination codons. Messenger RNAs that contain premature stop codons are degraded more rapidly than their wild-type counterparts, a phenomenon termed """"""""nonsense-mediated mRNA decay"""""""" (NMD). NMD occurs in all eukaryotes tested, and its action substantially impacts human genetic disease. Approximately one third of all sequenced point mutations that cause human genetic disease are nonsense or frameshift mutations, and most of these are known or predicted to elicit NMD of the mutant mRNA. We will study seven C. elegans genes required for NMD (smg-1 through smg-7) and will investigate their genetic and molecular properties. Our work falls into three broad categories: (1) investigating the molecular mechanisms of NMD;(2) investigating the relationship of NMD to RNA interference;and (3) investigating the biological function of NMD in wild-type animals.
Our Specific Aims are: 1. We will investigate the structure of mRNPs containing premature translation termination codons. Such mRNPs differ from those not containing premature stop codons by selectively binding the SMG-2 protein. 2. We will study the roles of SMG-2 phosphorylation and dephosphorylation in NMD. 3. We will investigate a recently discovered relationship between NMD and RNAi. We will test whether NMD contributes to gene silencing via endogenous RNAi. 4. We will identify most or all of the endogenous mRNA targets of NMD. Our long-range goals are to understand both the molecular mechanisms of mRNA turnover and the biological roles of NMD. Our experiments contribute to these goals by identifying proteins that are required for NMD, by describing their biochemical activities in vivo, and by investigating the role of NMD during normal growth and development.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM050933-16
Application #
8138350
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Bender, Michael T
Project Start
1994-05-01
Project End
2014-08-31
Budget Start
2011-09-01
Budget End
2014-08-31
Support Year
16
Fiscal Year
2011
Total Cost
$415,089
Indirect Cost
Name
University of Wisconsin Madison
Department
Genetics
Type
Schools of Earth Sciences/Natur
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
LeGendre, Jacqueline Baca; Campbell, Zachary T; Kroll-Conner, Peggy et al. (2013) RNA targets and specificity of Staufen, a double-stranded RNA-binding protein in Caenorhabditis elegans. J Biol Chem 288:2532-45
Izumi, Natsuko; Yamashita, Akio; Iwamatsu, Akihiro et al. (2010) AAA+ proteins RUVBL1 and RUVBL2 coordinate PIKK activity and function in nonsense-mediated mRNA decay. Sci Signal 3:ra27
Hubert, Amy; Anderson, Philip (2009) The C. elegans sex determination gene laf-1 encodes a putative DEAD-box RNA helicase. Dev Biol 330:358-67
Squirrell, Jayne M; Eggers, Zachary T; Luedke, Nancy et al. (2006) CAR-1, a protein that localizes with the mRNA decapping component DCAP-1, is required for cytokinesis and ER organization in Caenorhabditis elegans embryos. Mol Biol Cell 17:336-44