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PMID: 21726814 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

siRNA repositioning for guide strand selection by human Dicer complexes.

Molecular cell ·Vol. 43 ·No. 1 ·2011-07-08 ·Pages 110-21

Noland CL, Ma E, Doudna JA

Abstract

The human ribonuclease Dicer and its double-stranded RNA (dsRNA)-binding protein (dsRBP) partners TRBP and PACT play important roles in the biogenesis of regulatory RNAs. Following dicing, one dsRNA product strand is preferentially assembled into an RNA-induced silencing complex (RISC). The mechanism of strand selection in humans and the possible role of Dicer in this process remain unclear. Here we demonstrate that dsRNAs undergo significant repositioning within Dicer complexes following dicing. This repositioning enables directional binding of RNA duplexes, thereby biasing their orientation for guide strand selection according to the thermodynamic properties of the helix. Our findings indicate that Dicer is itself capable of sensing siRNA thermodynamic asymmetry regardless of the dsRBP to which it is bound. These results support a model in which Dicer employs two distinct RNA-binding sites-one for dsRNA processing and the other for sensing of siRNA thermodynamic asymmetry-during RISC loading in humans.

MeSH Terms
Binding Sites Humans Models, Genetic RNA, Small Interfering/chemistry,metabolism RNA-Induced Silencing Complex/chemistry,metabolism Ribonuclease III/physiology Thermodynamics
Chemicals
RNA, Small Interfering RNA-Induced Silencing Complex Ribonuclease III
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Noland Cameron L
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
Ma Enbo
Doudna Jennifer A
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44 references, click to expand
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Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-4164
DOI
10.1016/j.molcel.2011.05.028
Published
2011-07-08
Pages
110-21
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC3143821
Subset
IM
Grants
NIGMS NIH HHS · R01 GM073794 · United States
NIGMS NIH HHS · R01 GM073794-07 · United States
Howard Hughes Medical Institute · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2011.05.028 PMC Full Text View journal details
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