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Journal of Virology, September 2002, p. 9545-9550, Vol. 76, No. 18
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.18.9545-9550.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Interferon Regulatory Factor 3 Is a Cellular Partner of Rotavirus NSP1{dagger}

Joel W. Graff, Dana N. Mitzel, Carla M. Weisend, Michelle L. Flenniken, and Michele E. Hardy*

Veterinary Molecular Biology, Montana State University, Bozeman, Montana 59717

Received 11 March 2002/ Accepted 12 June 2002

The rotavirus nonstructural protein NSP1 is the least conserved protein in the rotavirus genome, and its function in the replication cycle is not known. We employed NSP1 as bait in the yeast two-hybrid interaction trap to identify candidate cellular partners of NSP1 that may provide clues to its function. Interferon regulatory factor 3 (IRF-3) was identified as an NSP1 interactor. NSP1 synthesized in rotavirus-infected cells bound IRF-3 in a glutathione S-transferase pull-down assay, indicating that the interaction was not unique to the two-hybrid system. NSP1 of murine rotavirus strain EW also interacted with IRF-3. NSP1 deletion and point mutants were constructed to map domains important in the interaction between NSP1 and IRF-3. The data suggest that a binding domain resides in the C terminus of NSP1 and that the N-terminal conserved zinc finger is important but not sufficient to mediate binding to IRF-3. We predict that a role for NSP1 in rotavirus-infected cells is to inhibit activation of IRF-3 and diminish the cellular interferon response.

* Corresponding author. Mailing address: Veterinary Molecular Biology, P.O. Box 173610, Montana State University, Bozeman, MT 59717. Phone: (406) 994-6378. Fax: (406) 994-4303. E-mail: mhardy{at}montana.edu.

{dagger} This article has been assigned journal number 2002-11 by the MAES.

Journal of Virology, September 2002, p. 9545-9550, Vol. 76, No. 18
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.18.9545-9550.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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