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Cited by 15DNA Repair MechanismsDNA and Nucleic Acid ChemistryGenomics and Chromatin Dynamics

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Structures of 9-1-1 DNA checkpoint clamp loading at gaps from start to finish and ramification on biology

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Zheng, Fengwei, Georgescu, Roxana E., Yao, Nina Y., O’Donnell, Michael E., Li, Huilin·2023·Cell Reports
DOI 10.1016/j.celrep.2023.112694PMC PMC10529453

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Abstract

Recent structural studies show the Rad24-RFC loads the 9-1-1 checkpoint clamp onto a recessed 5' end by binding the 5' DNA on Rad24 at an external surface site and threading the 3' ssDNA into the well-established internal chamber and into 9-1-1. We find here that Rad24-RFC loads 9-1-1 onto DNA gaps in preference to a recessed 5' DNA end, thus presumably leaving 9-1-1 on a 3' ss/ds DNA after Rad24-RFC ejects from the 5' gap end and may explain reports of 9-1-1 directly functioning in DNA repair with various TLS polymerases, in addition to signaling the ATR kinase. To gain a deeper understanding of 9-1-1 loading at gaps we report high-resolution structures of Rad24-RFC during loading of 9-1-1 onto 10-nt and 5-nt gapped DNAs. At a 10-nt gap we captured five Rad24-RFC-9-1-1 loading intermediates in which the 9-1-1 DNA entry gate varies from fully open to fully closed around DNA using ATPγS, supporting the emerging view that ATP hydrolysis is not needed for clamp opening/closing, but instead for dissociation of the loader from the clamp encircling DNA. The structure of Rad24-RFC-9-1-1 at a 5-nt gap shows a 180° axially rotated 3'-dsDNA which orients the template strand to bridge the 3'- and 5'- junctions with a minimum 5-nt ssDNA. The structures reveal a unique loop on Rad24 that limits the length of dsDNA in the inner chamber, and inability to melt DNA ends unlike RFC, thereby explaining Rad24-RFC's preference for a preexisting ssDNA gap and suggesting a direct role in gap repair in addition to its checkpoint role.

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Metadata

Authors
Zheng, F., Georgescu, R. E., Yao, N. Y., O’Donnell, M. E., & Li, H.
Affiliations
  • Van Andel Institute (US)
  • Rockefeller University (US)
Corresponding author
Li, Huilin
ORCID
Venue
Cell Reports
Publisher
Elsevier BV
Published
2023-07
Volume
42
Issue
7
Pages
112694
ISSN
2211-1247 (ISSN-L)
DOI
10.1016/j.celrep.2023.112694
PMC
PMC10529453
OpenAlex
W4382775869
Cited by
15
References
5
Citations by year
2026: 4 · 2025: 4 · 2024: 6 · 2023: 1
FWCI
1.34
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Reports (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Howard Hughes Medical Institute
  • NIGMS (GM115809, GM131754)
  • BCRF
  • National Institutes of Health
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
clamp loading, single-stranded DNA, DNA end resection, translesion synthesis, DNA replication checkpoint

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