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Journal of Virology, June 2002, p. 6244-6256, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6244-6256.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Gammaherpesvirus Lytic Gene Expression as Characterized by DNA Array

Joo Wook Ahn,1 Kenneth L. Powell,1 Paul Kellam,2 and Dagmar G. Alber1*

Wolfson Institute for Biomedical Research, University College London, London WC1E 6BT,1 Wohl Virion Centre, Department of Immunology and Molecular Pathology, Windeyer Institute, University College London, London W1T 4JF, United Kingdom2

Received 2 October 2001/ Accepted 5 March 2002

Gammaherpesviruses are associated with a number of diseases including lymphomas and other malignancies. Murine gammaherpesvirus 68 (MHV-68) constitutes the most amenable animal model for this family of pathogens. However experimental characterization of gammaherpesvirus gene expression, at either the protein or RNA level, lags behind that of other, better-studied alpha- and beta-herpesviruses. We have developed a cDNA array to globally characterize MHV-68 gene expression profiles, thus providing an experimental supplement to a genome that is chiefly annotated by homology. Viral genes started to be transcribed as early as 3 h postinfection (p.i.), and this was followed by a rapid escalation of gene expression that could be seen at 5 h p.i. Individual genes showed their own transcription profiles, and most genes were still being expressed at 18 h p.i. Open reading frames (ORFs) M3 (chemokine-binding protein), 52, and M9 (capsid protein) were particularly noticeable due to their very high levels of expression. Hierarchical cluster analysis of transcription profiles revealed four main groups of genes and allowed functional predictions to be made by comparing expression profiles of uncharacterized genes to those of genes of known function. Each gene was also categorized according to kinetic class by blocking de novo protein synthesis and viral DNA replication in vitro. One gene, ORF 73, was found to be expressed with {alpha}-kinetics, 30 genes were found to be expressed with ß-kinetics, and 42 genes were found to be expressed with {gamma}-kinetics. This fundamental characterization furthers the development of this model and provides an experimental basis for continued investigation of gammaherpesvirus pathology.

* Corresponding author. Mailing address: Wolfson Institute for Biomedical Research, The Cruciform Building, University College London, Gower St., London WC1E 6BT, United Kingdom. Phone: 44 20 7679 6718. Fax: 44 20 7813 2846. E-mail: d.alber{at}ucl.ac.uk.

Journal of Virology, June 2002, p. 6244-6256, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6244-6256.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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