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Biochemical and genetical analyses of the three mcm genes from the hyperthermophilic archaeon, Thermococcus kodakarensis
Author(s) -
Ishino Sonoko,
Fujino Seiji,
Tomita Hiroya,
Ogino Hiromi,
Takao Koichi,
Daiyasu Hiromi,
Kanai Tamotsu,
Atomi Haruyuki,
Ishino Yoshizumi
Publication year - 2011
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/j.1365-2443.2011.01562.x
Subject(s) - biology , helicase , minichromosome maintenance , thermococcus , hyperthermophile , replisome , dna replication , dnab helicase , microbiology and biotechnology , gene , origin recognition complex , pre replication complex , replication factor c , archaea , genetics , eukaryotic dna replication , rna
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.