This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Knez, J.
Right arrow Articles by Capone, J. P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Knez, J.
Right arrow Articles by Capone, J. P.

 Previous Article  |  Next Article 

Journal of Virology, March 2003, p. 2892-2902, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.2892-2902.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Single Amino Acid Substitution in Herpes Simplex Virus Type 1 VP16 Inhibits Binding to the Virion Host Shutoff Protein and Is Incompatible with Virus Growth

J. Knez, P. T. Bilan, and J. P. Capone*

Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada L8N 3Z5

Received 13 November 2002/ Accepted 20 November 2002

In addition to its well-established role in the activation of herpes simplex virus immediate-early gene transcription, VP16 interacts with and downregulates the function of the virion host shutoff protein (vhs), thereby attenuating vhs-mediated destruction of viral mRNAs and translational arrest at late times of infection. We have carried out two-hybrid analysis in vivo and protein-protein interaction assays in vitro to identify determinants in VP16 necessary for interaction with vhs. The minimal amino-terminal subfragment of VP16 capable of binding to vhs encompassed residues 1 to 345. Alteration of a single leucine at position 344 to alanine (L344A) in the context of the amino-terminal fragment of VP16 containing residues 1 to 404 was sufficient to abolish interaction with vhs in vitro and in vivo. Leu344 could be replaced with hydrophobic amino acids (Ile, Phe, Met, or Val) but not by Asn, Lys, or Pro, indicating that hydrophobicity is an important property of binding to vhs. VP16 harboring a loss-of-function mutation at L344 was not compromised in its ability to interact with host cell factor (HCF-1) or to activate transcription of viral immediate-early genes in transient-transfection assays. Virus complementation assays using the VP16-null virus 8MA and the VP16/vhs double-mutant virus 8MA{Delta}Sma showed that VP16(L344A) was able to complement the growth of 8MA{Delta}Sma but not 8MA. Thus, a single point mutation in VP16 uncouples binding to vhs from other functions of VP16 required for virus growth and indicates that direct physical association between VP16 and vhs is necessary to sustain a productive infection.

* Corresponding author. Mailing address: Department of Biochemistry, McMaster University, 1200 Main St. W., HSC 2E5C, Hamilton, Ontario L8N 3Z5, Canada. Phone: (905) 525-9140, ext. 22184. Fax: (905) 546-0800. E-mail: caponej{at}mcmaster.ca.

Journal of Virology, March 2003, p. 2892-2902, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.2892-2902.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Newcomb, W. W., Brown, J. C. (2009). Time-Dependent Transformation of the Herpesvirus Tegument. J. Virol. 83: 8082-8089 [Abstract] [Full Text]  
  • Kalamvoki, M., Qu, J., Roizman, B. (2008). Translocation and Colocalization of ICP4 and ICP0 in Cells Infected with Herpes Simplex Virus 1 Mutants Lacking Glycoprotein E, Glycoprotein I, or the Virion Host Shutoff Product of the UL41 Gene. J. Virol. 82: 1701-1713 [Abstract] [Full Text]  
  • Taddeo, B., Sciortino, M. T., Zhang, W., Roizman, B. (2007). Interaction of herpes simplex virus RNase with VP16 and VP22 is required for the accumulation of the protein but not for accumulation of mRNA. Proc. Natl. Acad. Sci. USA 104: 12163-12168 [Abstract] [Full Text]  
  • Yedowitz, J. C., Kotsakis, A., Schlegel, E. F. M., Blaho, J. A. (2005). Nuclear Localizations of the Herpes Simplex Virus Type 1 Tegument Proteins VP13/14, vhs, and VP16 Precede VP22-Dependent Microtubule Reorganization and VP22 Nuclear Import. J. Virol. 79: 4730-4743 [Abstract] [Full Text]  
  • Strand, S. S., Leib, D. A. (2004). Role of the VP16-Binding Domain of vhs in Viral Growth, Host Shutoff Activity, and Pathogenesis. J. Virol. 78: 13562-13572 [Abstract] [Full Text]  
  • Doepker, R. C., Hsu, W.-L., Saffran, H. A., Smiley, J. R. (2004). Herpes Simplex Virus Virion Host Shutoff Protein Is Stimulated by Translation Initiation Factors eIF4B and eIF4H. J. Virol. 78: 4684-4699 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»