Biochemical and genetical analyses of the three mcm genes from the hyperthermophilic archaeon, Thermococcus kodakarensis

Sonoko Ishino, Seiji Fujino, Hiroya Tomita, Hiromi Ogino, Koichi Takao, Hiromi Daiyasu, Tamotsu Kanai, Haruyuki Atomi, Yoshizumi Ishino

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)

Abstract

In eukaryotes, the replicative DNA helicase 'core' is the minichromosome maintenance (Mcm) complex (MCM), forming a heterohexameric complex consisting of six subunits (Mcm2-7). Recent studies showed that the CMG (Cdc45-MCM-GINS) complex is the actual helicase body in the replication fork progression complex. In Archaea, Thermococcus kodakarensis harbors three genes encoding the Mcm homologs on its genome, contrary to most archaea, which have only one homolog. It is thus, of high interest, whether and how these three Mcms share their functions in DNA metabolism in this hyperthermophile. Here, we report the biochemical properties of two of these proteins, TkoMcm1 and TkoMcm3. In addition, their physical and functional interactions with GINS, possibly an essential factor for the initiation and elongation process of DNA replication, are presented through in vitro ATPase and helicase assays, and an in vivo immunoprecipitation assay. Gene disruption and product quantification analyses suggested that TkoMcm3 is essential for cell growth and plays a key role as the main DNA helicase in DNA replication, whereas TkoMcm1 also shares some function in the cells.

Original languageEnglish
Pages (from-to)1176-1189
Number of pages14
JournalGenes to Cells
Volume16
Issue number12
DOIs
Publication statusPublished - Dec 1 2011

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

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