8B4K

Rfa1-N-terminal domain in complex with phosphorylated Ddc2

8B4K の概要

• エントリーDOI: 10.2210/pdb8b4k/pdb
• 分子名称: Replication factor A protein 1 (2 entities in total)
• 機能のキーワード: complex, ob-fold, replication protein a, dna damage response, replication, protein binding
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast)
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 45753.44

構造登録者

Yates, L.A.,Zhang, X. (登録日: 2022-09-20, 公開日: 2023-09-27)

主引用文献

Yates, L.A.,Tannous, E.A.,Morgan, R.M.,Burgers, P.M.,Zhang, X.
A DNA damage-induced phosphorylation circuit enhances Mec1 ATR Ddc2 ATRIP recruitment to Replication Protein A.
Proc.Natl.Acad.Sci.USA, 120:e2300150120-e2300150120, 2023

PubMed Abstract: The cell cycle checkpoint kinase Mec1 and its integral partner Ddc2 are vital for the DNA damage and replication stress response. Mec1-Ddc2 "senses" single-stranded DNA (ssDNA) by being recruited to the ssDNA binding Replication Protein A (RPA) via Ddc2. In this study, we show that a DNA damage-induced phosphorylation circuit modulates checkpoint recruitment and function. We demonstrate that Ddc2-RPA interactions modulate the association between RPA and ssDNA and that Rfa1-phosphorylation aids in the further recruitment of Mec1-Ddc2. We also uncover an underappreciated role for Ddc2 phosphorylation that enhances its recruitment to RPA-ssDNA that is important for the DNA damage checkpoint in yeast. The crystal structure of a phosphorylated Ddc2 peptide in complex with its RPA interaction domain provides molecular details of how checkpoint recruitment is enhanced, which involves Zn. Using electron microscopy and structural modeling approaches, we propose that Mec1-Ddc2 complexes can form higher order assemblies with RPA when Ddc2 is phosphorylated. Together, our results provide insight into Mec1 recruitment and suggest that formation of supramolecular complexes of RPA and Mec1-Ddc2, modulated by phosphorylation, would allow for rapid clustering of damage foci to promote checkpoint signaling.

PubMed: 36996117
DOI: 10.1073/pnas.2300150120

実験手法

X-RAY DIFFRACTION (1.55 Å)