De novo replication of the influenza virus RNA genome is regulated by DNA replicative helicase, MCM

EMBO J. 2007 Oct 31;26(21):4566-75. doi: 10.1038/sj.emboj.7601881. Epub 2007 Oct 11.

Abstract

By dissecting and reconstituting a cell-free influenza virus genome replication system, we have purified and identified the minichromosome maintenance (MCM) complex, which is thought to be a DNA replicative helicase, as one of the host factors that regulate the virus genome replication. MCM interacted with the PA subunit of the viral RNA-dependent RNA polymerase that is found to be involved in the replication genetically. The virus genome replication was decreased in MCM2 knockdown cells. The viral polymerase appeared to be a nonproductive complex, that is, it was capable of initiating replication but produced only abortive short RNA chains. MCM stimulated de novo-initiated replication reaction by stabilizing a replication complex during its transition from initiation to elongation. Based on the findings, including the result that the MCM-mediated RNA replication reaction was competed with exogenously added RNA, we propose that MCM functions as a scaffold between the nascent RNA chains and the viral polymerase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell-Free System
  • DNA Replication
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / genetics
  • Genome, Viral*
  • Hydrolysis
  • Minichromosome Maintenance 1 Protein / genetics
  • Models, Genetic
  • Nucleic Acid Conformation
  • Orthomyxoviridae / genetics*
  • RNA / chemistry
  • RNA, Viral / genetics*
  • Viral Proteins / chemistry
  • Virus Replication

Substances

  • Minichromosome Maintenance 1 Protein
  • RNA, Viral
  • Viral Proteins
  • RNA
  • DNA-Directed RNA Polymerases