Abstract

The broad objective of this project is to analyze at the molecular level the regulatory mechanisms of gene expression controlled by double-stranded RNA (dsRNA) in human cells. Molecules of dsRNA are either produced by viral infection or transposon activity, or they are expressed in a regulated manner from cellular genes that encode short inverted repeat sequences (microRNAs). Introduction of dsRNA into cells cognate to cellular genes has revolutionized cell biological studies enabling investigators to suppress their favorite gene for functional analysis and has opened the doors to new ways of gene-specific therapy.
The aim of this project is to identify and characterize the protein components involved in RNA silencing and to understand at molecular level how silencing-active ribonucleoprotein complexes are formed and exert their function. 1. Characterization of the human siRNA/miRNA-protein complexes. 1.1 Investigate the role of siRNA/miRNA-associated Argonaute protein members (elF2C family, HIWI, HILl) and define their interaction network with cellular components required for RNAi and miRNA-mediated gene regulation. 1.2 Reconstitute the sequence-specific endonuclease complex (RISC) from recombinant proteins and short duplex or single-stranded siRNA molecules. 2. Isolation of miRNP/target mRNA complexes and identification of the cellular mRNAs subjected to microRNA-mediated control.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM068476-04
Application #
7071819
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Rhoades, Marcus M
Project Start
2003-06-01
Project End
2008-05-31
Budget Start
2006-06-01
Budget End
2007-05-31
Support Year
4
Fiscal Year
2006
Total Cost
$293,046
Indirect Cost

  Institution

Name
Rockefeller University
Department
Genetics
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065

  Publications

Wang, Yanli; Juranek, Stefan; Li, Haitao et al. (2009) Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes. Nature 461:754-61
Hausser, Jean; Landthaler, Markus; Jaskiewicz, Lukasz et al. (2009) Relative contribution of sequence and structure features to the mRNA binding of Argonaute/EIF2C-miRNA complexes and the degradation of miRNA targets. Genome Res 19:2009-20
Chen, Po Yu; Weinmann, Lasse; Gaidatzis, Dimos et al. (2008) Strand-specific 5'-O-methylation of siRNA duplexes controls guide strand selection and targeting specificity. RNA 14:263-74
Landthaler, Markus; Gaidatzis, Dimos; Rothballer, Andrea et al. (2008) Molecular characterization of human Argonaute-containing ribonucleoprotein complexes and their bound target mRNAs. RNA 14:2580-96
Yuan, Yu-Ren; Pei, Yi; Chen, Hong-Ying et al. (2006) A potential protein-RNA recognition event along the RISC-loading pathway from the structure of A. aeolicus Argonaute with externally bound siRNA. Structure 14:1557-65
Patel, D J; Ma, J-B; Yuan, Y-R et al. (2006) Structural biology of RNA silencing and its functional implications. Cold Spring Harb Symp Quant Biol 71:81-93
Meister, Gunter; Landthaler, Markus; Peters, Lasse et al. (2005) Identification of novel argonaute-associated proteins. Curr Biol 15:2149-55
Yuan, Yu-Ren; Pei, Yi; Ma, Jin-Biao et al. (2005) Crystal structure of A. aeolicus argonaute, a site-specific DNA-guided endoribonuclease, provides insights into RISC-mediated mRNA cleavage. Mol Cell 19:405-19
Martinez, Javier; Tuschl, Thomas (2004) RISC is a 5' phosphomonoester-producing RNA endonuclease. Genes Dev 18:975-80
Meister, Gunter; Landthaler, Markus; Dorsett, Yair et al. (2004) Sequence-specific inhibition of microRNA- and siRNA-induced RNA silencing. RNA 10:544-50

Showing the most recent 10 out of 13 publications