7TJH

S. cerevisiae ORC bound to 84 bp ARS1 DNA and Cdc6 (state 1) with flexible Orc6 N-terminal domain

7TJH の概要

• エントリーDOI: 10.2210/pdb7tjh/pdb
• EMDBエントリー: 25924 25925 25926 25927 25928
• 分子名称: Origin recognition complex subunit 1, ADENOSINE-5'-TRIPHOSPHATE, MAGNESIUM ION, ... (12 entities in total)
• 機能のキーワード: initiator, helicase loader, aaa+ atpase, replication-dna complex, replication/dna
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast); 詳細
• タンパク質・核酸の鎖数: 9
• 化学式量合計: 527392.38

構造登録者

Schmidt, J.M.,Yang, R.,Kumar, A.,Hunker, O.,Bleichert, F. (登録日: 2022-01-16, 公開日: 2022-10-05, 最終更新日: 2025-06-04)

主引用文献

Schmidt, J.M.,Yang, R.,Kumar, A.,Hunker, O.,Seebacher, J.,Bleichert, F.
A mechanism of origin licensing control through autoinhibition of S. cerevisiae ORC·DNA·Cdc6.
Nat Commun, 13:1059-1059, 2022

PubMed Abstract: The coordinated action of multiple replicative helicase loading factors is needed for the licensing of replication origins prior to DNA replication. Binding of the Origin Recognition Complex (ORC) to DNA initiates the ATP-dependent recruitment of Cdc6, Cdt1 and Mcm2-7 loading, but the structural details for timely ATPase site regulation and for how loading can be impeded by inhibitory signals, such as cyclin-dependent kinase phosphorylation, are unknown. Using cryo-electron microscopy, we have determined several structures of S. cerevisiae ORC·DNA·Cdc6 intermediates at 2.5-2.7 Å resolution. These structures reveal distinct ring conformations of the initiator·co-loader assembly and inactive ATPase site configurations for ORC and Cdc6. The Orc6 N-terminal domain laterally engages the ORC·Cdc6 ring in a manner that is incompatible with productive Mcm2-7 docking, while deletion of this Orc6 region alleviates the CDK-mediated inhibition of Mcm7 recruitment. Our findings support a model in which Orc6 promotes the assembly of an autoinhibited ORC·DNA·Cdc6 intermediate to block origin licensing in response to CDK phosphorylation and to avert DNA re-replication.

PubMed: 35217664
DOI: 10.1038/s41467-022-28695-w

実験手法

ELECTRON MICROSCOPY (2.5 Å)