title: Mcm10 plays an essential role in origin DNA unwinding after loading of the CMG components.
abstract: The CMG complex composed of Mcm2-7, Cdc45 and GINS is postulated to be the eukaryotic replicative DNA helicase, whose activation requires sequential recruitment of replication proteins onto Mcm2-7. Current models suggest that Mcm10 is involved in assembly of the CMG complex, and in tethering of DNA polymerase α at replication forks. Here, we report that Mcm10 is required for origin DNA unwinding after association of the CMG components with replication origins in fission yeast. A combination of promoter shut-off and the auxin-inducible protein degradation (off-aid) system efficiently depleted cellular Mcm10 to <0.5% of the wild-type level. Depletion of Mcm10 did not affect origin loading of Mcm2-7, Cdc45 or GINS, but impaired recruitment of RPA and DNA polymerases. Mutations in a conserved zinc finger of Mcm10 abolished RPA loading after recruitment of Mcm10. These results show that Mcm10, together with the CMG components, plays a novel essential role in origin DNA unwinding through its zinc-finger function.
authors.affiliation: Department of Biological Sciences, Graduate School of Science, Osaka University, Osaka, Japan.
authors.forename: Mai
authors.initials: M
authors.lastname: Kanke
authors.forename: Yukako
authors.initials: Y
authors.lastname: Kodama
authors.forename: Tatsuro S
authors.initials: TS
authors.lastname: Takahashi
authors.forename: Takuro
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Hisao
authors.initials: H
authors.lastname: Masukata
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D004271:DNA, Fungal / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D015003:Yeasts
chemical_list: D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004271:DNA, Fungal; D005656:Fungal Proteins; D017493:Leukocyte Common Antigens