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

Three mcm genes from T. kodakarensis

descriptionPublicationkeyboard_double_arrow_right Article 17 Nov 2011 English Publisher:WileyJournal:Genes to Cells, volume 16, pages 1,176-1,189 (issn: 1356-9597,

Authors: Seiji Fujino; Sonoko Ishino; Hiromi Daiyasu; Koichi Takao; Hiroya Tomita; Hiromi Ogino; Yoshizumi Ishino; +2 Authors

doi: 10.1111/j.1365-2443.2011.01562.x

pmid: 22093166

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

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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.

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Keywords

DNA Replication, Hot Temperature, Chromosomal Proteins, Non-Histone, DNA Helicases, Recombinant Proteins, Polymerization, Isoenzymes, Thermococcus, Gene Knockdown Techniques, Escherichia coli, Immunoprecipitation, Anaerobiosis, Transformation, Bacterial, Cloning, Molecular, Plasmids, Protein Binding

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A Ctf4 trimer couples the CMG helicase to DNA polymerase α in the eukaryotic replisome
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