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PMID: 20184375 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Recognition of siRNA asymmetry by TAR RNA binding protein.

Biochemistry ·Vol. 49 ·No. 14 ·2010-04-13 ·Pages 3148-55

Gredell JA, Dittmer MJ, Wu M, Chan C, Walton SP

Abstract

The recognition of small interfering RNAs (siRNAs) by the RNA-induced silencing complex (RISC) and its precursor, the RISC loading complex (RLC), is a key step in the RNA interference pathway that controls the subsequent sequence-specific mRNA degradation. In Drosophila, selection of the guide strand has been shown to be mediated by RLC protein R2D2, which senses the relative hybridization stability between the two ends of the siRNA. A protein with similar function has yet to be conclusively identified in humans. We show here that human TAR RNA binding protein (TRBP) alone can bind siRNAs in vitro and sense their asymmetry. We also show that TRBP can bind 21-nucleotide single-stranded RNAs, though with far lower affinity than for double-stranded siRNA, and that TRBP cross-links preferentially to the 3'-ends of the guide strands of siRNAs. This suggests that TRBP binding depends both on the sequences of the siRNA strands and on the relative hybridization stability of the ends of the duplex. Together, these results demonstrate the importance of the siRNA-TRBP interaction in the selection of the siRNA guide strand in RNAi.

MeSH Terms
Cross-Linking Reagents/chemistry Humans Maltose-Binding Proteins Nucleic Acid Heteroduplexes/chemistry Periplasmic Binding Proteins/genetics Protein Binding RNA, Double-Stranded/chemistry RNA, Small Interfering/chemistry RNA-Binding Proteins/chemistry Recombinant Fusion Proteins/chemistry
Chemicals
Cross-Linking Reagents Maltose-Binding Proteins Nucleic Acid Heteroduplexes Periplasmic Binding Proteins RNA, Double-Stranded RNA, Small Interfering RNA-Binding Proteins Recombinant Fusion Proteins trans-activation responsive RNA-binding protein
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gredell Joseph A
Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, Michigan 48824-1226, USA.
Dittmer Michael J
Wu Ming
Chan Christina
Walton S Patrick
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Article Info
Journal
Biochemistry
Abbr.
Biochemistry
ISSN
1520-4995
DOI
10.1021/bi902189s
Published
2010-04-13
Pages
3148-55
Language
English
Region
United States
NLM ID
0370623
PMCID
PMC2851484
Subset
IM
Grants
NCRR NIH HHS · R21 RR024439-02 · United States
NIGMS NIH HHS · GM079688 · United States
NIGMS NIH HHS · R01 GM079688 · United States
NCI NIH HHS · CA126136 · United States
NCI NIH HHS · R21 CA126136-02 · United States
NCI NIH HHS · R21 CA126136 · United States
NCRR NIH HHS · RR024439 · United States
NIGMS NIH HHS · R01 GM079688-04 · United States
NCRR NIH HHS · R21 RR024439 · United States
External Links
PubMed source DOI: 10.1021/bi902189s PMC Full Text View journal details
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