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Journal of Virology, August 2005, p. 9566-9571, Vol. 79, No. 15
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.15.9566-9571.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Determination of Interactions between Tegument Proteins of Herpes Simplex Virus Type 1

Valerio Vittone, Eve Diefenbach, Damian Triffett,{dagger} Mark W. Douglas, Anthony L. Cunningham, and Russell J. Diefenbach*

Centre For Virus Research, The Westmead Millennium Institute, The University of Sydney and Westmead Hospital, Westmead, NSW 2145, Australia

Received 7 December 2004/ Accepted 20 April 2005

The aim of this study was to elucidate protein-protein interactions between tegument proteins of herpes simplex virus type 1 (HSV-1). To do so, we have cloned and expressed in the LexA yeast (Saccharomyces cerevisiae) two-hybrid system, 13 of the 21 currently known tegument proteins of HSV-1. These included the tegument proteins essential for replication in cell lines, UL17, UL36, UL37, UL48, and UL49, and the nonessential tegument proteins US11, UL11, UL14, UL16, UL21, UL41, UL46, and UL47. A total of 104 combinations were screened in the yeast two-hybrid assay, with 9 interactions identified. These included: UL11-UL16, UL36-UL37, UL36-UL48, UL46-UL48, UL47-UL48, and UL48-UL49. The remaining interactions consisted of self-associations that were observed for US11, UL37, and UL49. The interactions UL36-UL37, UL36-UL48, UL37-UL37, UL46-UL48, and UL47-UL48 have not been previously reported for HSV-1. The interaction of UL46-UL48 was verified using an in vitro pull-down assay. The interactions of UL36-UL37 and UL37-UL37 were verified with a coimmunoprecipitation assay. Knowledge of HSV-1 tegument protein-protein interactions will provide insights into the pathways of tegument assembly, and the identified interactions are potential targets for new antiviral drugs.

* Corresponding author. Mailing address: Centre for Virus Research, Westmead Millennium Institute, P.O. Box 412, Westmead 2145 NSW, Australia. Phone: 61 2 9845 9111. Fax: 61 2 9845 9103. E-mail: russell_diefenbach{at}wmi.usyd.edu.au.

{dagger} Present address: Molecular Genetics Group, John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia.

Journal of Virology, August 2005, p. 9566-9571, Vol. 79, No. 15
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.15.9566-9571.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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