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PMID: 16862146 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

The endocytic pathway mediates cell entry of dsRNA to induce RNAi silencing.

Nature cell biology ·Vol. 8 ·No. 8 ·2006-08-00 ·Pages 793-802

Saleh MC, van Rij RP, Hekele A, Gillis A, Foley E, O'Farrell PH, Andino R

Abstract

Many metazoan cells can take up exogenous double-stranded (ds) RNA and use it to initiate an RNA silencing response, however, the mechanism for this uptake is ill-defined. Here, we identify the pathway for dsRNA uptake in Drosophila melanogaster S2 cells. Biochemical and cell biological analyses, and a genome-wide screen for components of the dsRNA-uptake machinery, indicated that dsRNA is taken up by an active process involving receptor-mediated endocytosis. Pharmacological inhibition of endocytic pathways disrupted exogenous dsRNA entry and the induction of gene silencing. This dsRNA uptake mechanism seems to be evolutionarily conserved, as knockdown of orthologues in Caenorhabditis elegans inactivated the RNA interference response in worms. Thus, this entry pathway is required for systemic RNA silencing in whole organisms. In Drosophila cells, pharmacological evidence suggests that dsRNA entry is mediated by pattern-recognition receptors. The possible role of these receptors in dsRNA entry may link RNA interference (RNAi) silencing to other innate immune responses.

MeSH Terms
Animals Caenorhabditis elegans/drug effects,genetics,metabolism Cell Line DNA/genetics,metabolism Drosophila/cytology,drug effects,metabolism Endocytosis/drug effects,physiology Luciferases/genetics,metabolism Microscopy, Fluorescence Polysaccharides/pharmacology RNA Interference/physiology RNA, Double-Stranded/chemistry,genetics,metabolism Signal Transduction/drug effects,physiology
Chemicals
Polysaccharides RNA, Double-Stranded DNA fucoidan Luciferases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Saleh Maria-Carla
Department of Microbiology and Immunology, University of California San Francisco, CA 94143-2280, USA.
van Rij Ronald P
Hekele Armin
Gillis Amethyst
Foley Edan
O'Farrell Patrick H
Andino Raul
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Article Info
Journal
Nature cell biology
Abbr.
Nat Cell Biol
ISSN
1465-7392
DOI
10.1038/ncb1439
Published
2006-08-00
Epub
2006-00-23
Pages
793-802
Language
English
Region
England
NLM ID
100890575
PMCID
PMC2731564
Subset
IM
Grants
NIAID NIH HHS · N01AI40085 · United States
NIAID NIH HHS · R01 AI060102 · United States
NIAID NIH HHS · R01 AI060102-07 · United States
External Links
PubMed source DOI: 10.1038/ncb1439 PMC Full Text View journal details
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