Cited by 363DNA Repair MechanismsCancer-related Molecular PathwaysDNA and Nucleic Acid Chemistry

Biochemical Characterization of DNA Damage Checkpoint Complexes: Clamp Loader and Clamp Complexes with Specificity for 5′ Recessed DNA

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Ellison, Viola, Stillman, Bruce·2003·PLoS Biology

Abstract

The cellular pathways involved in maintaining genome stability halt cell cycle progression in the presence of DNA damage or incomplete replication. Proteins required for this pathway include Rad17, Rad9, Hus1, Rad1, and Rfc-2, Rfc-3, Rfc-4, and Rfc-5. The heteropentamer replication factor C (RFC) loads during DNA replication the homotrimer proliferating cell nuclear antigen (PCNA) polymerase clamp onto DNA. Sequence similarities suggest the biochemical functions of an RSR (Rad17-Rfc2-Rfc3-Rfc4-Rfc5) complex and an RHR heterotrimer (Rad1-Hus1-Rad9) may be similar to that of RFC and PCNA, respectively. RSR purified from human cells loads RHR onto DNA in an ATP-, replication protein A-, and DNA structure-dependent manner. Interestingly, RSR and RFC differed in their ATPase activities and displayed distinct DNA substrate specificities. RSR preferred DNA substrates possessing 5' recessed ends whereas RFC preferred 3' recessed end DNA substrates. Characterization of the biochemical loading reaction executed by the checkpoint clamp loader RSR suggests new insights into the mechanisms underlying recognition of damage-induced DNA structures and signaling to cell cycle controls. The observation that RSR loads its clamp onto a 5' recessed end supports a potential role for RHR and RSR in diverse DNA metabolism, such as stalled DNA replication forks, recombination-linked DNA repair, and telomere maintenance, among other processes.

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Authors
Ellison, V., & Stillman, B.
Affiliations
Cold Spring Harbor Laboratory (US)
Corresponding author
Stillman, Bruce
Venue
PLoS Biology
Published
2003-11-17
Volume
1
Issue
2
Pages
e33
ISSN
  • 1544-9173 (ISSN-L)
  • 1545-7885
OpenAlex
W2101942126
Cited by
363
References
80
Citations by year
2026: 7 · 2025: 5 · 2024: 11 · 2023: 6 · 2022: 13 · 2021: 9 · 2020: 15 · 2019: 6 · 2018: 10 · 2017: 12 · 2016: 14 · 2015: 12 · 2014: 21 · 2013: 22 · 2012: 11
FWCI
8.52
Type
journal article
Language
en
Open access
Locations
  • PLoS Biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, other-oa)
  • PubMed (publishedVersion)
  • Cold Spring Harbor Laboratory Institutional Repository (Cold Spring Harbor Laboratory) (acceptedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
National Cancer Institute (CA13106, P01CA013106)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
Keywords
DNA damage checkpoint, clamp loader, replication factor C, proliferating cell nuclear antigen, DNA loading, ATPase activity, genome stability, cell cycle arrest, DNA damage recognition, stalled replication forks, telomere maintenance
MeSH terms
Adenosine Triphosphate, DNA, DNA Damage, DNA Replication, Escherichia coli, Exonucleases, Humans, Models, Biological, Molecular Sequence Data, Protein Binding, Recombination, Genetic, Saccharomyces cerevisiae, Schizosaccharomyces, Substrate Specificity, Telomere, Protein Structure, Secondary, Cell Cycle Proteins, Proliferating Cell Nuclear Antigen, Dimerization, Schizosaccharomyces pombe Proteins, Cell Line, Tumor, Immunoprecipitation, Replication Protein C

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