Mcm10 plays an essential role in origin DNA unwinding after loading of the CMG components

Mai Kanke, Yukako Kodama, Tatsuro S. Takahashi, Takuro Nakagawa, Hisao Masukata

Research output: Contribution to journalArticle

71 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2182-2194
Number of pages13
JournalEMBO Journal
Volume31
Issue number9
DOIs
Publication statusPublished - May 2 2012
Externally publishedYes

Fingerprint

Zinc Fingers
DNA-Directed DNA Polymerase
Zinc
DNA Helicases
Replication Origin
Indoleacetic Acids
Schizosaccharomyces
DNA
Yeast
Proteolysis
Proteins
Chemical activation
Association reactions
Degradation
Mutation

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Mcm10 plays an essential role in origin DNA unwinding after loading of the CMG components. / Kanke, Mai; Kodama, Yukako; Takahashi, Tatsuro S.; Nakagawa, Takuro; Masukata, Hisao.

In: EMBO Journal, Vol. 31, No. 9, 02.05.2012, p. 2182-2194.

Research output: Contribution to journalArticle

Kanke, Mai ; Kodama, Yukako ; Takahashi, Tatsuro S. ; Nakagawa, Takuro ; Masukata, Hisao. / Mcm10 plays an essential role in origin DNA unwinding after loading of the CMG components. In: EMBO Journal. 2012 ; Vol. 31, No. 9. pp. 2182-2194.
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