De novo replication of the influenza virus RNA genome is regulated by DNA replicative helicase, MCM.
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.
Kawaguchi
Atsushi
A
Department of Infection Biology, Graduate School of Comprehensive Human Sciences and Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba, Japan.
Nagata
Kyosuke
K
D002474:Cell-Free System; D004261:DNA Replication; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics; D016679:Genome, Viral; D006868:Hydrolysis; D026341:Minichromosome Maintenance 1 Protein / Q000235:genetics; D008957:Models, Genetic; D009690:Nucleic Acid Conformation; D009975:Orthomyxoviridae / Q000235:genetics*; D012313:RNA / Q000737:chemistry; D012367:RNA, Viral / Q000235:genetics*; D014764:Viral Proteins / Q000737:chemistry; D014779:Virus Replication
D026341:Minichromosome Maintenance 1 Protein; D012367:RNA, Viral; D014764:Viral Proteins; D012313:RNA; D012321:DNA-Directed RNA Polymerases