7P5Z

Structure of a DNA-loaded MCM double hexamer engaged with the Dbf4-dependent kinase

これはPDB形式変換不可エントリーです。

7P5Z の概要

• エントリーDOI: 10.2210/pdb7p5z/pdb
• 関連するPDBエントリー: 7P30
• EMDBエントリー: 13176 13211
• 分子名称: DNA replication licensing factor MCM2, DNA (53-MER), ADENOSINE-5'-TRIPHOSPHATE, ... (14 entities in total)
• 機能のキーワード: kinase, mcm2-7 helicase, nucleoprotein complex, dna replication, replication
• 由来する生物種: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); 詳細
• タンパク質・核酸の鎖数: 16
• 化学式量合計: 1401822.80

構造登録者

Greiwe, J.F.,Miller, T.C.R.,Martino, F.,Costa, A. (登録日: 2021-07-15, 公開日: 2022-02-02, 最終更新日: 2024-07-17)

主引用文献

Greiwe, J.F.,Miller, T.C.R.,Locke, J.,Martino, F.,Howell, S.,Schreiber, A.,Nans, A.,Diffley, J.F.X.,Costa, A.
Structural mechanism for the selective phosphorylation of DNA-loaded MCM double hexamers by the Dbf4-dependent kinase.
Nat.Struct.Mol.Biol., 29:10-20, 2022

PubMed Abstract: Loading of the eukaryotic replicative helicase onto replication origins involves two MCM hexamers forming a double hexamer (DH) around duplex DNA. During S phase, helicase activation requires MCM phosphorylation by Dbf4-dependent kinase (DDK), comprising Cdc7 and Dbf4. DDK selectively phosphorylates loaded DHs, but how such fidelity is achieved is unknown. Here, we determine the cryogenic electron microscopy structure of Saccharomyces cerevisiae DDK in the act of phosphorylating a DH. DDK docks onto one MCM ring and phosphorylates the opposed ring. Truncation of the Dbf4 docking domain abrogates DH phosphorylation, yet Cdc7 kinase activity is unaffected. Late origin firing is blocked in response to DNA damage via Dbf4 phosphorylation by the Rad53 checkpoint kinase. DDK phosphorylation by Rad53 impairs DH phosphorylation by blockage of DDK binding to DHs, and also interferes with the Cdc7 active site. Our results explain the structural basis and regulation of the selective phosphorylation of DNA-loaded MCM DHs, which supports bidirectional replication.

PubMed: 34963704
DOI: 10.1038/s41594-021-00698-z

実験手法

ELECTRON MICROSCOPY (3.3 Å)