Premium
Mcm10 plays an essential role in origin DNA unwinding after loading of the CMG components
Author(s) -
Kanke Mai,
Kodama Yukako,
Takahashi Tatsuro S,
Nakagawa Takuro,
Masukata Hisao
Publication year - 2012
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1038/emboj.2012.68
Subject(s) - biology , dna , genetics , microbiology and biotechnology , computational biology , evolutionary biology
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.