7STB

Closed state of Rad24-RFC:9-1-1 bound to a 5' ss/dsDNA junction

Summary for 7STB

• Entry DOI: 10.2210/pdb7stb/pdb
• EMDB information: 25423
• Descriptor: Checkpoint protein RAD24, DNA (5'-D(P*CP*GP*CP*TP*CP*CP*TP*TP*CP*CP*TP*GP*AP*CP*TP*CP*GP*TP*CP*C)-3'), PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER, ... (13 entities in total)
• Functional Keywords: dna damage, dna replication, dna sliding clamp, replication, replication-dna complex, replication/dna
• Biological source: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); More
• Total number of polymer chains: 10
• Total formula weight: 431363.78

Authors

Castaneda, J.C.,Schrecker, M.,Remus, D.,Hite, R.K. (deposition date: 2021-11-12, release date: 2022-03-23, Last modification date: 2024-06-05)

Primary citation

Castaneda, J.C.,Schrecker, M.,Remus, D.,Hite, R.K.
Mechanisms of loading and release of the 9-1-1 checkpoint clamp.
Nat.Struct.Mol.Biol., 29:369-375, 2022

PubMed Abstract: Single-stranded or double-stranded DNA junctions with recessed 5' ends serve as loading sites for the checkpoint clamp, 9-1-1, which mediates activation of the apical checkpoint kinase, ATR. However, the basis for 9-1-1's recruitment to 5' junctions is unclear. Here, we present structures of the yeast checkpoint clamp loader, Rad24-replication factor C (RFC), in complex with 9-1-1 and a 5' junction and in a post-ATP-hydrolysis state. Unexpectedly, 9-1-1 adopts both closed and planar open states in the presence of Rad24-RFC and DNA. Moreover, Rad24-RFC associates with the DNA junction in the opposite orientation of processivity clamp loaders with Rad24 exclusively coordinating the double-stranded region. ATP hydrolysis stimulates conformational changes in Rad24-RFC, leading to disengagement of DNA-loaded 9-1-1. Together, these structures explain 9-1-1's recruitment to 5' junctions and reveal new principles of sliding clamp loading.

PubMed: 35314831
DOI: 10.1038/s41594-022-00741-7

Experimental method

ELECTRON MICROSCOPY (2.72 Å)