Regulation of viral intermediate gene expression by the vaccinia virus B1 protein kinase

G. R. Kovacs, Nikos Vasilakis, B. Moss

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The BI gene of vaccinia virus encodes a serine/threonine protein kinase that is expressed early after infection. Under nonpermissive conditions, temperature-sensitive mutants (ts2 and ts25) that map to B1 fail to efficiently replicate viral DNA. Our goal was to extend studies on the function of B1 by determining if the kinase is required for intermediate or late gene expression, two events that ordinarily depend on viral DNA replication. First, we established that early viral gene expression occurred at the nonpermissive temperature. By using a transfection procedure that circumvents the viral DNA replication requirement, we found that reporter genes regulated by an intermediate promoter were transcribed only under conditions permissive for expression of active B1. To assay late gene expression, the T7 RNA polymerase gene was inserted into the genome of ts25 to form ts25/T7. A DNA replication-independent late gene transcription system was established by cotransfecting plasmids containing T7 promoter-driven late gene transcription factors and a late promoter reporter gene into ts25/T7-infected cells. Late genes, unlike intermediate genes, were expressed at the nonpermissive temperature. Last, we showed that overexpression of B1 stimulated intermediate but inhibited late gene expression in cells infected with wild-type virus.

Original languageEnglish (US)
Pages (from-to)4048-4055
Number of pages8
JournalJournal of Virology
Volume75
Issue number9
DOIs
StatePublished - 2001
Externally publishedYes

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Viral Gene Expression Regulation
Vaccinia virus
protein kinases
Viral DNA
DNA Replication
Genetic Transcription
Gene Expression
gene expression
DNA replication
Reporter Genes
Genes
Temperature
genes
promoter regions
reporter genes
Viral Genes
Protein-Serine-Threonine Kinases
Transfection
temperature
Plasmids

ASJC Scopus subject areas

  • Immunology

Cite this

Regulation of viral intermediate gene expression by the vaccinia virus B1 protein kinase. / Kovacs, G. R.; Vasilakis, Nikos; Moss, B.

In: Journal of Virology, Vol. 75, No. 9, 2001, p. 4048-4055.

Research output: Contribution to journalArticle

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