Cell cycle stage-specific roles of Rad18 in tolerance and repair of oxidative DNA damage
Open accessAbstract
The E3 ubiquitin ligase Rad18 mediates tolerance of replication fork-stalling bulky DNA lesions, but whether Rad18 mediates tolerance of bulky DNA lesions acquired outside S-phase is unclear. Using synchronized cultures of primary human cells, we defined cell cycle stage-specific contributions of Rad18 to genome maintenance in response to ultraviolet C (UVC) and H(2)O(2)-induced DNA damage. UVC and H(2)O(2) treatments both induced Rad18-mediated proliferating cell nuclear antigen mono-ubiquitination during G(0), G(1) and S-phase. Rad18 was important for repressing H(2)O(2)-induced (but not ultraviolet-induced) double strand break (DSB) accumulation and ATM S1981 phosphorylation only during G(1), indicating a specific role for Rad18 in processing of oxidative DNA lesions outside S-phase. However, H(2)O(2)-induced DSB formation in Rad18-depleted G1 cells was not associated with increased genotoxin sensitivity, indicating that back-up DSB repair mechanisms compensate for Rad18 deficiency. Indeed, in DNA LigIV-deficient cells Rad18-depletion conferred H(2)O(2)-sensitivity, demonstrating functional redundancy between Rad18 and non-homologous end joining for tolerance of oxidative DNA damage acquired during G(1). In contrast with G(1)-synchronized cultures, S-phase cells were H(2)O(2)-sensitive following Rad18-depletion. We conclude that although Rad18 pathway activation by oxidative lesions is not restricted to S-phase, Rad18-mediated trans-lesion synthesis by Polη is dispensable for damage-tolerance in G(1) (because of back-up non-homologous end joining-mediated DSB repair), yet Rad18 is necessary for damage tolerance during S-phase.
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Hydrogen peroxide triggers ATR-Chk1 checkpoint signaling in human dermal fibroblasts HDF cells and in Xenopus egg extracts.
“More recently, it has been demonstrated that hydrogen peroxide triggers ATR-Chk1 checkpoint signaling in human dermal fibroblasts HDF cells [137] and in Xenopus egg extracts [86].”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [137]
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- Authors
- Yang, Y., Durando, M., Smith-Roe, S. L., Sproul, C., Greenwalt, A. M., Kaufmann, W., Oh, S., Hendrickson, E. A., & Vaziri, C.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- Corresponding author
- Yang, Yang
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2013-01-07
- Volume
- 41
- Issue
- 4
- Pages
- 2296-2312
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC3575850
- OpenAlex
- W2027848959
- Cited by
- 33
- References
- 64
- Citations by year
- 2026: 3 · 2025: 2 · 2023: 2 · 2022: 2 · 2021: 3 · 2020: 2 · 2019: 2 · 2018: 4 · 2017: 4 · 2016: 2 · 2015: 3 · 2014: 4
- FWCI
- 1.34
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Nucleic Acids Research (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- UNC Libraries
- Funders
- National Institutes of Health
- University of North Carolina at Chapel Hill
- National Cancer Institute (R01CA154461)
- National Institute of General Medical Sciences (R01GM088351)
- National Institute of Environmental Health Sciences (ES09558, ES016280, R01ES016280, F30ES019449, R01ES009558, P30ES010126)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
- Keywords
- RAD18, E3 ubiquitin ligase, oxidative DNA damage, DNA damage tolerance, replication fork stalling, DNA double-strand break repair, non-homologous end joining, DNA ligase IV, translesion synthesis, pol, ultraviolet C, hydrogen peroxide, genome maintenance
- MeSH terms
- Y-Family DNA Polymerases, Cell Cycle, Cells, Cultured, DNA Damage, DNA-Directed DNA Polymerase, DNA Repair, DNA-Binding Proteins, Humans, Hydrogen Peroxide, Oxidation-Reduction, G1 Phase, S Phase, Proliferating Cell Nuclear Antigen, Ubiquitin-Protein Ligases, Replication Protein A, DNA Breaks, Double-Stranded, Ubiquitination
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