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Journal of Virology, September 2006, p. 8664-8675, Vol. 80, No. 17
0022-538X/06/$08.00+0     doi:10.1128/JVI.00498-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Characterization of a UL49-Null Mutant: VP22 of Herpes Simplex Virus Type 1 Facilitates Viral Spread in Cultured Cells and the Mouse Cornea

Carol Duffy,1 Jennifer H. LaVail,2 Andrew N. Tauscher,2 Elizabeth G. Wills,1 John A. Blaho,3 and Joel D. Baines1*

Department of Microbiology and Immunology, Cornell University, Ithaca, New York 14853,1 Departments of Anatomy and Ophthalmology, University of California San Francisco, San Francisco, California 94143-0452,2 Department of Microbiology, Mount Sinai School of Medicine, New York, New York 100293

Received 9 March 2006/ Accepted 12 June 2006

Herpes simplex virus type 1 (HSV-1) virions, like those of all herpesviruses, contain a proteinaceous layer termed the tegument that lies between the nucleocapsid and viral envelope. The HSV-1 tegument is composed of at least 20 different viral proteins of various stoichiometries. VP22, the product of the UL49 gene, is one of the most abundant tegument proteins and is conserved among the alphaherpesviruses. Although a number of interesting biological properties have been attributed to VP22, its role in HSV-1 infection is not well understood. In the present study we have generated both a UL49-null virus and its genetic repair and characterized their growth in both cultured cells and the mouse cornea. While single-step growth analyses indicated that VP22 is dispensable for virus replication at high multiplicities of infection (MOIs), analyses of plaque morphology and intra- and extracellular multistep growth identified a role for VP22 in viral spread during HSV-1 infection at low MOIs. Specifically, VP22 was not required for either virion infectivity or cell-cell spread but was required for accumulation of extracellular virus to wild-type levels. We found that the absence of VP22 also affected virion composition. Intracellular virions generated by the UL49-null virus contained reduced amounts of ICP0 and glycoproteins E and D compared to those generated by the wild-type and UL49-repaired viruses. In addition, viral spread in the mouse cornea was significantly reduced upon infection with the UL49-null virus compared to infection with the wild-type and UL49-repaired viruses, identifying a role for VP22 in viral spread in vivo as well as in vitro.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Cornell University, C5143 Veterinary Education Center, Ithaca, NY 14853. Phone: (607) 253-3391. Fax: (607) 253-3384. E-mail: jdb11{at}cornell.edu.

Journal of Virology, September 2006, p. 8664-8675, Vol. 80, No. 17
0022-538X/06/$08.00+0     doi:10.1128/JVI.00498-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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