title: Biochemical and genetical analyses of the three mcm genes from the hyperthermophilic archaeon, Thermococcus kodakarensis.
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.
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki, Higashi-ku, Fukuoka, Japan.
authors.forename: Sonoko
authors.initials: S
authors.lastname: Ishino
authors.forename: Seiji
authors.initials: S
authors.lastname: Fujino
authors.forename: Hiroya
authors.initials: H
authors.lastname: Tomita
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ogino
authors.forename: Koichi
authors.initials: K
authors.lastname: Takao
authors.forename: Hiromi
authors.initials: H
authors.lastname: Daiyasu
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Kanai
authors.forename: Haruyuki
authors.initials: H
authors.lastname: Atomi
authors.forename: Yoshizumi
authors.initials: Y
authors.lastname: Ishino
mesh_terms: D000693:Anaerobiosis; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004261:DNA Replication / Q000235:genetics*; D004926:Escherichia coli; D055785:Gene Knockdown Techniques; D006358:Hot Temperature; D047468:Immunoprecipitation; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids; D058105:Polymerization; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019712:Thermococcus / Q000235:genetics / Q000378:metabolism*; D014169:Transformation, Bacterial
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D007527:Isoenzymes; D011994:Recombinant Proteins; D004265:DNA Helicases