7QHS

S. cerevisiae CMGE nucleating origin DNA melting

これはPDB形式変換不可エントリーです。

7QHS の概要

• エントリーDOI: 10.2210/pdb7qhs/pdb
• EMDBエントリー: 13978 13988
• 分子名称: DNA replication licensing factor MCM2, Cell division control protein 45, DNA polymerase epsilon subunit B, ... (19 entities in total)
• 機能のキーワード: dna replication, helicase, initiation, dna origin, replication
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast); 詳細
• タンパク質・核酸の鎖数: 15
• 化学式量合計: 1148414.47

構造登録者

Lewis, J.S.,Sousa, J.S.,Costa, A. (登録日: 2021-12-14, 公開日: 2022-06-15, 最終更新日: 2024-07-17)

主引用文献

Lewis, J.S.,Gross, M.H.,Sousa, J.,Henrikus, S.S.,Greiwe, J.F.,Nans, A.,Diffley, J.F.X.,Costa, A.
Mechanism of replication origin melting nucleated by CMG helicase assembly.
Nature, 606:1007-1014, 2022

PubMed Abstract: The activation of eukaryotic origins of replication occurs in temporally separated steps to ensure that chromosomes are copied only once per cell cycle. First, the MCM helicase is loaded onto duplex DNA as an inactive double hexamer. Activation occurs after the recruitment of a set of firing factors that assemble two Cdc45-MCM-GINS (CMG) holo-helicases. CMG formation leads to the underwinding of DNA on the path to the establishment of the replication fork, but whether DNA becomes melted at this stage is unknown. Here we use cryo-electron microscopy to image ATP-dependent CMG assembly on a chromatinized origin, reconstituted in vitro with purified yeast proteins. We find that CMG formation disrupts the double hexamer interface and thereby exposes duplex DNA in between the two CMGs. The two helicases remain tethered, which gives rise to a splayed dimer, with implications for origin activation and replisome integrity. Inside each MCM ring, the double helix becomes untwisted and base pairing is broken. This comes as the result of ATP-triggered conformational changes in MCM that involve DNA stretching and protein-mediated stabilization of three orphan bases. Mcm2 pore-loop residues that engage DNA in our structure are dispensable for double hexamer loading and CMG formation, but are essential to untwist the DNA and promote replication. Our results explain how ATP binding nucleates origin DNA melting by the CMG and maintains replisome stability at initiation.

PubMed: 35705812
DOI: 10.1038/s41586-022-04829-4

実験手法

ELECTRON MICROSCOPY (3.3 Å)