7STE

Rad24-RFC ADP state

Summary for 7STE

• Entry DOI: 10.2210/pdb7ste/pdb
• EMDB information: 25426
• Descriptor: Replication factor C subunit 4, Replication factor C subunit 3, Replication factor C subunit 2, ... (8 entities in total)
• Functional Keywords: dna damage, dna replication, dna sliding clamp, replication
• Biological source: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); More
• Total number of polymer chains: 5
• Total formula weight: 236481.63

Authors

Castaneda, J.C.,Schrecker, M.,Remus, D.,Hite, R.K. (deposition date: 2021-11-12, release date: 2022-04-06, 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 (2.73 Å)