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PMID: 15556947 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

ADAR1 RNA deaminase limits short interfering RNA efficacy in mammalian cells.

The Journal of biological chemistry ·Vol. 280 ·No. 5 ·2005-02-04 ·Pages 3946-53

Yang W, Wang Q, Howell KL, Lee JT, Cho DS, Murray JM, Nishikura K

Abstract

Double-stranded RNA induces the homology-dependent degradation of cognate mRNA in the cytoplasm via RNA interference (RNAi) but also is a target for adenosine-to-inosine (A-to-I) RNA editing by adenosine deaminases acting on RNA (ADARs). An interaction between the RNAi and the RNA editing pathways in Caenorhabditis elegans has been suggested recently, but the precise mode of interaction remains to be established. In addition, it is unclear whether this interaction is possible in mammalian cells with their somewhat different RNAi pathways. Here we show that ADAR1 and ADAR2, but not ADAR3, avidly bind short interfering RNA (siRNA) without RNA editing. In particular, the cytoplasmic full-length isoform of ADAR1 has the highest affinity among known ADARs, with a subnanomolar dissociation constant. Gene silencing by siRNA is significantly more effective in mouse fibroblasts homozygous for an ADAR1 null mutation than in wild-type cells. In addition, suppression of RNAi effects are detected in fibroblast cells overexpressing functional ADAR1 but not when overexpressing mutant ADAR1 lacking double-stranded RNA-binding domains. These results identify ADAR1 as a cellular factor that limits the efficacy of siRNA in mammalian cells.

MeSH Terms
Adenosine Deaminase/genetics,metabolism Animals Cells, Cultured Fibroblasts/cytology Gene Expression Gene Silencing/physiology Mammals Mice Protein Binding/genetics RNA Editing/physiology RNA, Small Interfering/metabolism RNA-Binding Proteins
Chemicals
RNA, Small Interfering RNA-Binding Proteins ADARB1 protein, human Adenosine Deaminase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Yang Weidong
The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
Wang Qingde
Howell Kelly L
Lee Joshua T
Cho Dan-Sung C
Murray John M
Nishikura Kazuko
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M407876200
Published
2005-02-04
Epub
2004-00-19
Pages
3946-53
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2947832
Subset
IM
Grants
NIGMS NIH HHS · R01 GM040536 · United States
NHLBI NIH HHS · R01 HL070045 · United States
NIGMS NIH HHS · R01 GM040536-15 · United States
NCI NIH HHS · P01 CA072765 · United States
NHLBI NIH HHS · R01 HL070045-02 · United States
NCI NIH HHS · P01 CA072765-05S20002 · United States
External Links
PubMed source DOI: 10.1074/jbc.M407876200 PMC Full Text View journal details
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