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Journal of Virology, May 2001, p. 4068-4079, Vol. 75, No. 9
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.9.4068-4079.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The SH Integral Membrane Protein of the Paramyxovirus Simian Virus 5 Is Required To Block Apoptosis in MDBK Cells

Biao He,1 Grace Y. Lin,2 Joan E. Durbin,3 Russell K. Durbin,3 and Robert A. Lamb1,2,*

Howard Hughes Medical Institute1 and Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 60208-3500,2 and Children's Research Institute, Children's Hospital, and Division of Pathology, Department of Pediatrics, College of Medicine and Public Health, The Ohio State University, Columbus, Ohio 432053

Received 7 November 2000/Accepted 5 February 2001

In some cell types the paramyxovirus simian virus 5 (SV5) causes little cytopathic effect (CPE) and infection continues productively for long periods of time; e.g., SV5 can be produced from MDBK cells for up to 40 days with little CPE. SV5 differs from most paramyxoviruses in that it encodes a small (44-amino-acid) hydrophobic integral membrane protein (SH). When MDBK cells were infected with a recombinant SV5 containing a deletion of the SH gene (rSV5Delta SH), the MDBK cells exhibited an increase in CPE compared to cells infected with wild-type SV5 (recovered from cDNA; rSV5). The increased CPE correlated with an increase in apoptosis in rSV5Delta SH-infected cells over mock-infected and rSV5-infected cells when assayed for annexin V binding, DNA content (propidium iodide staining), and DNA fragmentation (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling assay). In rSV5Delta SH-infected MDBK cells an increase in caspase-2 and caspase-3 activities was observed. By using peptide inhibitors of individual caspases it was found that caspase-2 and caspase-3 were activated separately in rSV5Delta SH-infected cells. Expression of caspase-2 and -3 in rSV5Delta SH-infected MDBK cells appeared not to require STAT1 protein, as STAT1 protein could not be detected in SV5-infected MDBK cells. When mutant mice homologous for a targeted disruption of STAT1 were used as a model animal system and infected with the viruses it was found that rSV5Delta SH caused less mortality than wild-type rSV5, consistent with the notion of clearance of apoptotic cells in a host species.

* Corresponding author. Dept. of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, 2153 North Campus Dr., Evanston, IL 60208-3500. Phone: (847) 491-5433. Fax: (847) 491-2467. E-mail: ralamb{at}northwestern.edu.

Journal of Virology, May 2001, p. 4068-4079, Vol. 75, No. 9
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.9.4068-4079.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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