Replication Factor C Is a More Effective Proliferating Cell Nuclear Antigen (PCNA) Opener than the Checkpoint Clamp Loader, Rad24-RFC
Open accessAbstract
Clamp loaders from all domains of life load clamps onto DNA. The clamp tethers DNA polymerases to DNA to increase the processivity of synthesis as well as the efficiency of replication. Here, we investigated proliferating cell nuclear antigen (PCNA) binding and opening by the Saccharomyces cerevisiae clamp loader, replication factor C (RFC), and the DNA damage checkpoint clamp loader, Rad24-RFC, using two separate fluorescence intensity-based assays. Analysis of PCNA opening by RFC revealed a two-step reaction in which RFC binds PCNA before opening PCNA rather than capturing clamps that have transiently and spontaneously opened in solution. The affinity of RFC for PCNA is about an order of magnitude lower in the absence of ATP than in its presence. The affinity of Rad24-RFC for PCNA in the presence of ATP is about an order magnitude weaker than that of RFC for PCNA, similar to the RFC-PCNA interaction in the absence of ATP. Importantly, fewer open clamp loader-clamp complexes are formed when PCNA is bound by Rad24-RFC than when bound by RFC.
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The intact RFC complex and PCNA are essential replication factors needed for loading the replicative polymerase onto DNA.
“Both the intact RFC complex and PCNA are essential replication factors and are needed for loading the replicative polymerase onto DNA 116, 117,”
From Xeroderma Pigmentosa Group A (XPA), Nucleotide Excision Repair and Regulation by ATR in Response to Ultraviolet Irradiation (2017) · cites this paper as [116]
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- Authors
- Thompson, J. A., Marzahn, M. R., O'Donnell, M., & Bloom, L. B.
- Affiliations
- University of Florida (US)
- Corresponding author
- Bloom, Linda B.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2012-01
- Volume
- 287
- Issue
- 3
- Pages
- 2203-2209
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC3265898
- OpenAlex
- W1970916666
- Cited by
- 37
- References
- 32
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 1 · 2023: 2 · 2022: 1 · 2020: 3 · 2019: 4 · 2018: 4 · 2017: 3 · 2016: 2 · 2015: 3 · 2014: 5 · 2013: 3 · 2012: 2
- FWCI
- 1.30
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of General Medical Sciences (R01GM082849, GM082849, R37GM038839, R01GM038839, GM038839)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- replication factor C, PCNA, proliferating cell nuclear antigen, clamp loader, DNA damage checkpoint, clamp loading, DNA polymerase processivity, ATP-dependent binding, Saccharomyces cerevisiae, DNA replication
- MeSH terms
- Protein Binding, Saccharomyces cerevisiae, Cell Cycle Proteins, Proliferating Cell Nuclear Antigen, Multiprotein Complexes, Intracellular Signaling Peptides and Proteins, Cell Proliferation, Replication Protein C
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