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Journal of Virology, June 2005, p. 7371-7379, Vol. 79, No. 12
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.12.7371-7379.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

A Virus-Encoded Inhibitor That Blocks RNA Interference in Mammalian Cells

Christopher S. Sullivan* and Don Ganem

Howard Hughes Medical Institute, Departments of Microbiology & Immunology and Medicine, G. W. Hooper Research Foundation, University of California Medical Center, San Francisco, California 94143-0414

Received 5 January 2005/ Accepted 14 February 2005

Nodamura virus (NoV) is a small RNA virus that is infectious for insect and mammalian hosts. We have developed a highly sensitive assay of RNA interference (RNAi) in mammalian cells that shows that the NoV B2 protein functions as an inhibitor of RNAi triggered by either short hairpin RNAs or small interfering RNAs. In the cell, NoV B2 binds to pre-Dicer substrate RNA and RNA-induced silencing complex (RISC)-processed RNAs and inhibits the Dicer cleavage reaction and, potentially, one or more post-Dicer activities. In vitro, NoV B2 inhibits Dicer-mediated RNA cleavage in the absence of any other host factors and specifically binds double-stranded RNAs corresponding in structure to Dicer substrates and products. Its abilities to bind to Dicer precursor and post-Dicer RISC-processed RNAs suggest a mechanism of inhibition that is unique among known viral inhibitors of RNAi.

* Corresponding author. Mailing address: G. W. Hooper Research Foundation, University of California, San Francisco, 513 Parnassus Ave., HSW 1501, Box 0552, San Francisco, CA 94143-0552. Phone: (415) 502-5669. Fax: (415) 476-0939. E-mail: csulliv1{at}itsa.ucsf.edu.

Journal of Virology, June 2005, p. 7371-7379, Vol. 79, No. 12
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.12.7371-7379.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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