Cryo-EM structure of a helicase loading intermediate containing ORC-Cdc6-Cdt1-MCM2-7 bound to DNA

Jingchuan Sun, Cecile Evrin, Stefan A. Samel, Alejandra Fernández-Cid, Alberto Riera, Hironori Kawakami, Bruce Stillman, Christian Speck, Huilin Li

研究成果: Contribution to journalArticle査読

103 被引用数 (Scopus)

抄録

In eukaryotes, the Cdt1-bound replicative helicase core MCM2-7 is loaded onto DNA by the ORC-Cdc6 ATPase to form a prereplicative complex (pre-RC) with an MCM2-7 double hexamer encircling DNA. Using purified components in the presence of ATP-γS, we have captured in vitro an intermediate in pre-RC assembly that contains a complex between the ORC-Cdc6 and Cdt1-MCM2-7 heteroheptamers called the OCCM. Cryo-EM studies of this 14-subunit complex reveal that the two separate heptameric complexes are engaged extensively, with the ORC-Cdc6 N-terminal AAA+ domains latching onto the C-terminal AAA+ motor domains of the MCM2-7 hexamer. The conformation of ORC-Cdc6 undergoes a concerted change into a right-handed spiral with helical symmetry that is identical to that of the DNA double helix. The resulting ORC-Cdc6 helicase loader shows a notable structural similarity to the replication factor C clamp loader, suggesting a conserved mechanism of action.

本文言語英語
ページ(範囲)944-951
ページ数8
ジャーナルNature Structural and Molecular Biology
20
8
DOI
出版ステータス出版済み - 8 2013

All Science Journal Classification (ASJC) codes

  • 構造生物学
  • 分子生物学

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