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Journal of Virology, April 2000, p. 2981-2989, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Reovirus-Induced Apoptosis Requires Activation of Transcription Factor NF-kappa B

Jodi L. Connolly,1,2 Steven E. Rodgers,1,2 Penny Clarke,3 Dean W. Ballard,1 Lawrence D. Kerr,1,4 Kenneth L. Tyler,3,5,6,7,8 and Terence S. Dermody1,2,9,*

Departments of Microbiology and Immunology,1 Cell Biology,4 and Pediatrics9 and Elizabeth B. Lamb Center for Pediatric Research,2 Vanderbilt University School of Medicine, Nashville, Tennessee 37232, and Departments of Neurology,3 Medicine,5 Microbiology,6 and Immunology,7 University of Colorado Health Sciences Center, and Neurology Service, Denver Veterans Affairs Medical Center,8 Denver, Colorado 80220

Received 5 October 1999/Accepted 29 December 1999

Reovirus infection induces apoptosis in cultured cells and in vivo. To identify host cell factors that mediate this response, we investigated whether reovirus infection alters the activation state of the transcription factor nuclear factor kappa B (NF-kappa B). As determined in electrophoretic mobility shift assays, reovirus infection of HeLa cells leads to nuclear translocation of NF-kappa B complexes containing Rel family members p50 and p65. Reovirus-induced activation of NF-kappa B DNA-binding activity correlated with the onset of NF-kappa B-directed transcription in reporter gene assays. Three independent lines of evidence indicate that this functional form of NF-kappa B is required for reovirus-induced apoptosis. First, treatment of reovirus-infected HeLa cells with a proteasome inhibitor prevents NF-kappa B activation following infection and substantially diminishes reovirus-induced apoptosis. Second, transient expression of a dominant-negative form of Ikappa B that constitutively represses NF-kappa B activation significantly reduces levels of apoptosis triggered by reovirus infection. Third, mutant cell lines deficient for either the p50 or p65 subunits of NF-kappa B are resistant to reovirus-induced apoptosis compared with cells expressing an intact NF-kappa B signaling pathway. These findings indicate that NF-kappa B plays a significant role in the mechanism by which reovirus induces apoptosis in susceptible host cells.

* Corresponding author. Mailing address: Lamb Center for Pediatric Research, D7235 MCN, Vanderbilt University School of Medicine, Nashville, TN 37232. Phone: (615) 343-9943. Fax: (615) 343-9723. E-mail: terry.dermody{at}mcmail.vanderbilt.edu.

Journal of Virology, April 2000, p. 2981-2989, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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