Temperature-sensitive mutant in the vaccinia virus E6 protein produce virions that are transcriptionally inactive

Virology. 2010 Apr 10;399(2):221-30. doi: 10.1016/j.virol.2010.01.010. Epub 2010 Feb 8.

Abstract

The vaccinia virus E6R gene encodes a late protein that is packaged into virion cores. A temperature-sensitive mutant was used to study the role of this protein in viral replicative cycle. Cts52 has a P226L missense mutation in the E6R gene, shows a two-log reduction in plaque formation, but displays normal patterns of gene expression, late protein processing and DNA replication during infection. Mutant virions produced at 40 degrees C were similar in their morphology to wt virions grown at 40 degrees C. The particle to infectivity ratio was 50 times higher in purified Cts52 grown at 40 degrees C when compared to the mutant grown at permissive temperature. In vitro characterization of Cts-52 particles grown at 40 degrees C revealed no differences in protein composition or in DNA content and the mutant virions could bind and enter cells. However, core particles prepared from Cts52 grown at 40 degrees C failed to transcribe in vitro. Our results show that E6 in the virion has either a direct or an indirect role in viral transcription.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • DNA, Viral / biosynthesis
  • DNA, Viral / genetics
  • Hot Temperature*
  • Mutation
  • Transcription, Genetic*
  • Vaccinia virus / genetics
  • Vaccinia virus / metabolism
  • Vaccinia virus / physiology*
  • Viral Core Proteins / genetics
  • Viral Core Proteins / metabolism*
  • Viral Plaque Assay
  • Virion / genetics
  • Virion / metabolism
  • Virion / physiology
  • Virus Attachment
  • Virus Internalization
  • Virus Replication*

Substances

  • DNA, Viral
  • Viral Core Proteins