H2AX phosphorylation within the G 1 phase after UV irradiation depends on nucleotide excision repair and not DNA double-strand breaks
Open accessAbstract
The variant histone H2AX is phosphorylated in response to UV irradiation of primary human fibroblasts in a complex fashion that is radically different from that commonly reported after DNA double-strand breaks. H2AX phosphorylation after exposure to ionizing radiation produces foci, which are detectable by immunofluorescence microscopy and have been adopted as clear and consistent quantitative markers for DNA double-strand breaks. Here we show that in contrast to ionizing radiation, UV irradiation mainly induces H2AX phosphorylation as a diffuse, even, pan-nuclear staining. UV induced pan-nuclear phosphorylation of H2AX is present in all phases of the cell cycle and is highest in S phase. H2AX phosphorylation in G 1 cells depends on nucleotide excision repair factors that may expose the S-139 site to kinase activity, is not due to DNA double-strand breaks, and plays a larger role in UV-induced signal transduction than previously realized.
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H2AX is phosphorylated both in G1 and S phases by ATR in response to UV irradiation.
“H2AX is phosphorylated both in G1 and S phases by ATR in response to UV irradiation [26;32–38].”
From ATR- and ATM-Mediated DNA Damage Response Is Dependent on Excision Repair Assembly during G1 but Not in S Phase of Cell Cycle (2016) · cites this paper as [33]
ATR is predominantly responsible for the DNA damage response upon UVR exposure.
“Previous studies support that ATR is predominantly responsible for DDR upon UVR exposure”
From ATR- and ATM-Mediated DNA Damage Response Is Dependent on Excision Repair Assembly during G1 but Not in S Phase of Cell Cycle (2016) · cites this paper as [33]
In G1, checkpoint activation is due to ssDNA and not from DSBs.
“Previous studies showed that in G1, checkpoint activation is due to the ssDNA and not from DSBs [33;46;49]”
From ATR- and ATM-Mediated DNA Damage Response Is Dependent on Excision Repair Assembly during G1 but Not in S Phase of Cell Cycle (2016) · cites this paper as [33]
Several studies have reported that NER factor-mediated lesion processing might be required for γH2AX foci formation.
“However, several studies reported that lesion processing by NER factors might be an essential step in γH2AX foci formation [35-39].”
From NER initiation factors, DDB2 and XPC, regulate UV radiation response by recruiting ATR and ATM kinases to DNA damage sites (2013) · cites this paper as [36]
In human XP cells, lack of NER-dependent processing prevents UV-induced checkpoint activity in non-cycling fibroblasts, and XP cells are deficient in both repair and G1/G2/M UV-induced checkpoint.
“Extension of this kind of analysis to human cells derived from XP patients confirmed that lack of NER-dependent processing prevented UV-induced checkpoint activity in non cycling fibroblasts, revealing that XP cells are not only deficient in repairing UV lesions, but they are also deficient in the G1 and G2/M UV-induced checkpoint [56,57].”
From Mind the gap: keeping UV lesions in check (2011) · cites this paper as [57]
Outside S phase and in non-cycling cells, UV lesions alone cannot activate the checkpoint; NER plays a major role in triggering the checkpoint response, though contrasting results exist.
“Outside of S phase and in non cycling cells the situation is quite different. Recent analysis showed that UV lesions themselves cannot activate the checkpoint and NER plays a major role in triggering the checkpoint response, although contrasting results have also been reported [52–57].”
From Mind the gap: keeping UV lesions in check (2011) · cites this paper as [57]
DDB2 and XPC bring ATM and ATR to UV-damaged sites and enhance ATM-Chk2 and ATR-Chk1 checkpoint signaling.
“DDB2 and XPC recruit ATM and ATR to UV-damaged sites and promote ATM-Chk2 and ATR-Chk1 checkpoint signaling.”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [202]
In G1 cells, UV-induced phosphorylation of H2AX at serine 139 requires both XPA and XPC.
“UV-induced H2AX phosphorylation at Serine 139 in G1 cells requires both XPA and XPC [202]”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [202]
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- Authors
- Marti, T. M., Hefner, E., Feeney, L., Natale, V., & Cleaver, J. E.
- Affiliations
- University of California, San Francisco (US)
- Corresponding author
- Cleaver, James E.
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2006-06-27
- Volume
- 103
- Issue
- 26
- Pages
- 9891-9896
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC1502549
- OpenAlex
- W2008162481
- Cited by
- 359
- References
- 34
- Citations by year
- 2026: 6 · 2025: 12 · 2024: 6 · 2023: 13 · 2022: 13 · 2021: 16 · 2020: 9 · 2019: 21 · 2018: 17 · 2017: 9 · 2016: 25 · 2015: 16 · 2014: 22 · 2013: 17 · 2012: 22
- FWCI
- 10.34
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation
- Iris O'Brien Foundation
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (102308)
- University of California, San Francisco
- National Cancer Institute (P30-CA82103, P30CA082103)
- National Institute of Environmental Health Sciences (1 R01 ES8061, R01ES008061)
- National Institute of Arthritis and Musculoskeletal and Skin Diseases (P01AR050440, T32AR007175)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
- Keywords
- H2AX phosphorylation, nucleotide excision repair, UV irradiation, G1 phase, DNA double-strand breaks, histone variants, DNA damage response, cell cycle, signal transduction, human fibroblasts
- MeSH terms
- Cell Nucleus, Cells, Cultured, DNA Damage, DNA Repair, DNA-Binding Proteins, Fibroblasts, Histones, Humans, Phosphorylation, Ultraviolet Rays, G1 Phase, Xeroderma Pigmentosum Group A Protein
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