7TFL

Atomic model of S. cerevisiae clamp loader RFC bound to DNA

7TFL の概要

• エントリーDOI: 10.2210/pdb7tfl/pdb
• EMDBエントリー: 25872 25873 25874 25875 25876
• 分子名称: Replication factor C subunit 1, ADENOSINE-5'-DIPHOSPHATE, Replication factor C subunit 4, ... (10 entities in total)
• 機能のキーワード: dna replication, dna damage repair, rfc loader, pcna clamp, dna polymerase processivity factor, dna binding protein-dna complex, dna binding protein/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: 7
• 化学式量合計: 270260.07

構造登録者

Zheng, F.,Georgescu, R.,Yao, Y.N.,O'Donnell, M.E.,Li, H. (登録日: 2022-01-06, 公開日: 2022-11-16, 最終更新日: 2025-05-28)

主引用文献

Zheng, F.,Georgescu, R.,Yao, N.Y.,Li, H.,O'Donnell, M.E.
Cryo-EM structures reveal that RFC recognizes both the 3'- and 5'-DNA ends to load PCNA onto gaps for DNA repair.
Elife, 11:-, 2022

PubMed Abstract: RFC uses ATP to assemble PCNA onto primed sites for replicative DNA polymerases δ and ε. The RFC pentamer forms a central chamber that binds 3' ss/ds DNA junctions to load PCNA onto DNA during replication. We show here five structures that identify a second DNA binding site in RFC that binds a 5' duplex. This 5' DNA site is located between the N-terminal BRCT domain and AAA+ module of the large Rfc1 subunit. Our structures reveal ideal binding to a 7-nt gap, which includes 2 bp unwound by the clamp loader. Biochemical studies show enhanced binding to 5 and 10 nt gaps, consistent with the structural results. Because both 3' and 5' ends are present at a ssDNA gap, we propose that the 5' site facilitates RFC's PCNA loading activity at a DNA damage-induced gap to recruit gap-filling polymerases. These findings are consistent with genetic studies showing that base excision repair of gaps greater than 1 base requires PCNA and involves the 5' DNA binding domain of Rfc1. We further observe that a 5' end facilitates PCNA loading at an RPA coated 30-nt gap, suggesting a potential role of the RFC 5'-DNA site in lagging strand DNA synthesis.

PubMed: 35829698
DOI: 10.7554/eLife.77469

実験手法

ELECTRON MICROSCOPY (3.33 Å)