JVI Figure table search 04
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
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 Yang, W. C.
Right arrow Articles by Triezenberg, S. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yang, W. C.
Right arrow Articles by Triezenberg, S. J.

 Previous Article  |  Next Article 

Journal of Virology, December 2002, p. 12758-12774, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12758-12774.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

General and Specific Alterations in Programming of Global Viral Gene Expression during Infection by VP16 Activation-Deficient Mutants of Herpes Simplex Virus Type 1

William C. Yang,1 G. V. Devi-Rao,2 Peter Ghazal,3 Edward K. Wagner,2* and Steven J. Triezenberg1*

Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824-1319,1 Department of Molecular Biology and Biochemistry and Center for Virus Research, University of California, Irvine, California 92717,2 Genomic Technology and Informatics Centre, University of Edinburgh, Summerhall, Edinburgh EH9 1QH, United Kingdom3

Received 12 June 2002/ Accepted 13 September 2002

During productive infection by herpes simplex virus 1 (HSV-1), viral gene expression occurs in a temporally regulated cascade in which transcription of the viral immediate-early (IE) genes is strongly stimulated by the virion protein VP16. We have employed an oligonucleotide microarray to examine the effect of VP16 mutations on the overall pattern of viral gene expression following infection of HeLa cells. This microarray detects essentially all HSV-1 transcripts with relative and absolute levels correlating well with known kinetics of expression. This analysis revealed that deletion of the VP16 activation domain sharply reduced overall viral gene expression; moreover, the pattern of this reduced expression varied greatly from the pattern of a wild-type (wt) infection. However, when this mutant virus was delivered at a high multiplicity of infection or in the presence of the cellular stress inducer hexamethylene bisacetamide, expression was largely restored to the wt levels and pattern. Infection with virions that deliver wt VP16 protein at the start of infection but synthesize only truncated VP16 resulted in a normal kinetic cascade. This suggests that newly synthesized VP16 does not play a significant role in the expression of later classes of transcripts. The VP16 activation domain comprises two subregions. Deletion of the C-terminal subregion resulted in minimal changes in the level and profile of gene expression compared to a normal (wt) cascade. In contrast, deletion of the N-terminal subregion reduced the overall expression levels and skewed the relative levels of IE transcripts but did not significantly alter the kinetic pattern of early and late transcript expression. We conclude that the general activation of IE gene transcription by VP16, but not the specific ratios of IE transcripts, is necessary for the subsequent ordered expression of viral genes. Moreover, this report establishes the feasibility of microarray analysis for globally assessing viral gene expression programs as a function of the conditions of infection.

* Corresponding author. Mailing address for S. J. Triezenberg: Department of Biochemistry and Molecular Biology, 510 Biochemistry Building, Michigan State University, East Lansing, MI 48824-1319. Phone: (517) 353-7120. Fax: (517) 353-9334. E-mail: triezenb{at}msu.edu. Mailing address for E. K. Wagner: Department of Molecular Biology and Biochemistry and Center for Virus Research, University of California, Irvine, CA 92717-3900. Phone: (949) 824-5370. Fax: (949) 824-8551. E-mail: ewagner{at}uci.edu.

Journal of Virology, December 2002, p. 12758-12774, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12758-12774.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Mol. Cell. Biol. Microbiol. Mol. Biol. Rev.
Clin. Vaccine Immunol. ALL ASM JOURNALS

Copyright © 2002 by the American Society for Microbiology. All rights reserved.