Identification of the putative RNA polymerase of Cryphonectria hypovirus in a solubilized replication complex

T. Fahima, Y. Wu, L. Zhang, N. K. Van Alfen

Research output: Contribution to journalComment/Debate

Abstract

Hypovirulence of the pathogenic fungus Cryphonectria parasitica, caused by the unencapsidated viral double-stranded RNA of Cryphonectria hypovirus (CHV1), provides a means for biological control of chestnut blight. We report here the isolation of a replication complex of the virus solubilized from host membranes. The conserved regions of the putative RNA polymerase encoded by strain CHV1-713 were cloned and expressed, and the recombinant protein was purified and used to produce polyclonal antibodies. The CHV1 replication complex was solubilized from a membrane fraction of CHV1-infected C. parasitica hyphae. Antibodies raised against the putative viral polymerase reacted on a Western immunoblot with an 87-kDa polypeptide of the replication complex but not with comparable preparations from an isogenic uninfected strain. Analysis of the polypeptide composition of the complex by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining revealed a number of other polypeptides along with the double-stranded RNA of the virus. We conclude that this 87-kDa polypeptide is the putative RNA polymerase encoded on open reading frame B of CHV1.

Original languageEnglish
Pages (from-to)6116-6119
Number of pages4
JournalJournal of Virology
Volume68
Issue number9
DOIs
StatePublished - Sep 1994

ASJC Scopus subject areas

  • Insect Science
  • Virology
  • Microbiology
  • Immunology

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