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Journal of Virology, July 2003, p. 7945-7956, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.7945-7956.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Ebola Virus VP35 Protein Inhibits Activation of Interferon Regulatory Factor 3

Christopher F. Basler,1* Andrea Mikulasova,1 Luis Martinez-Sobrido,1 Jason Paragas,2 Elke Mühlberger,3 Mike Bray,2,{dagger} Hans-Dieter Klenk,3 Peter Palese,1 and Adolfo García-Sastre1

Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029,1 Virology Division, US Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Frederick, Maryland 21702-5011,2 Institut für Virologie, Philipps-Universität Marburg, D-35037 Marburg, Germany3

Received 21 March 2003/ Accepted 1 May 2003

The Ebola virus VP35 protein was previously found to act as an interferon (IFN) antagonist which could complement growth of influenza delNS1 virus, a mutant influenza virus lacking the influenza virus IFN antagonist protein, NS1. The Ebola virus VP35 could also prevent the virus- or double-stranded RNA-mediated transcriptional activation of both the beta IFN (IFN-ß) promoter and the IFN-stimulated ISG54 promoter (C. Basler et al., Proc. Natl. Acad. Sci. USA 97:12289-12294, 2000). We now show that VP35 inhibits virus infection-induced transcriptional activation of IFN regulatory factor 3 (IRF-3)-responsive mammalian promoters and that VP35 does not block signaling from the IFN-{alpha}/ß receptor. The ability of VP35 to inhibit this virus-induced transcription correlates with its ability to block activation of IRF-3, a cellular transcription factor of central importance in initiating the host cell IFN response. We demonstrate that VP35 blocks the Sendai virus-induced activation of two promoters which can be directly activated by IRF-3, namely, the ISG54 promoter and the ISG56 promoter. Further, expression of VP35 prevents the IRF-3-dependent activation of the IFN-{alpha}4 promoter in response to viral infection. The inhibition of IRF-3 appears to occur through an inhibition of IRF-3 phosphorylation. VP35 blocks virus-induced IRF-3 phosphorylation and subsequent IRF-3 dimerization and nuclear translocation. Consistent with these observations, Ebola virus infection of Vero cells activated neither transcription from the ISG54 promoter nor nuclear accumulation of IRF-3. These data suggest that in Ebola virus-infected cells, VP35 inhibits the induction of antiviral genes, including the IFN-ß gene, by blocking IRF-3 activation.

* Corresponding author. Mailing address: Dept. of Microbiology, Box 1124, Mount Sinai School of Medicine, 1 Gustave L. Levy Pl., New York, NY 10029. Phone: (212) 241-5923. Fax: (212) 534-1684. E-mail: chris.basler{at}mssm.edu.

{dagger} Present address: National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, MD 20892.

Journal of Virology, July 2003, p. 7945-7956, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.7945-7956.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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