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Journal of Virology, July 2000, p. 6287-6299, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Evidence that Herpes Simplex Virus VP16 Is Required for Viral Egress Downstream of the Initial Envelopment Event

Karen L. Mossman,1 Richard Sherburne,1 Carol Lavery,2 Joanne Duncan,2 and James R. Smiley1,2,*

Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada T6G 2H7,1 and Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada L8N 3Z52

Received 25 February 2000/Accepted 19 April 2000

During infection with herpes simplex virus type 1 (HSV-1), VP16 serves multiple functions, including transcriptional activation of viral immediate early genes and downregulation of the virion host shutoff protein vhs. Furthermore, VP16 has been shown to be involved in some aspect of virus assembly and/or maturation. Experiments with a VP16 null virus, 8MA, suggested that VP16 plays a direct role in virion assembly, since removal of VP16 from the HSV-1 genome results in reduced levels of encapsidated DNA and a failure to produce extracellular enveloped particles. However, VP16 null mutants display a severe translational arrest due to unrestrained vhs activity, thus complicating interpretation of these data. We examine here the role of VP16 in virion assembly and egress in the context of a vhs null background, using the virus 8MA/Delta Sma (VP16- vhs-). Comparison of 8MA and 8MA/Delta Sma with respect to viral DNA accumulation and encapsidation and accumulation of the major capsid protein, VP5, revealed that the 8MA lethal phenotype is only partially due to uncontrolled vhs activity, indicating that VP16 is required in HSV-1 virion formation. Electron microscopy confirmed these results and further showed that VP16 is required for HSV-1 egress beyond the perinuclear space. In addition, we describe the isolation and characterization of an 8MA derivative capable of propagation on Vero cells, due to second site mutations in the vhs and UL53 (gK) genes. Taken together, these results show that VP16 is required for viral egress downstream of the initial envelopment step and further underscore the importance of VP16 in controlling vhs activity within an infected cell.

* Corresponding author. Mailing address: Department of Medical Microbiology and Immunology, 1-41, Medical Sciences Bldg., University of Alberta, Edmonton, Alberta, Canada T6G 2H7. Phone: (780) 492-2308. Fax: (780) 492-7521. E-mail: jim.smiley{at}ualberta.ca.

Journal of Virology, July 2000, p. 6287-6299, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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