Perturbed gap-filling synthesis in nucleotide excision repair causes histone H2AX phosphorylation in human quiescent cells
Open accessAbstract
Human histone H2AX is rapidly phosphorylated on serine 139 in response to DNA double-strand breaks and plays a crucial role in tethering the factors involved in DNA repair and damage signaling. Replication stress caused by hydroxyurea or UV also initiates H2AX phosphorylation in S-phase cells, although UV-induced H2AX phosphorylation in non-cycling cells has recently been observed. Here we study the UV-induced H2AX phosphorylation in human primary fibroblasts under growth-arrested conditions. This reaction absolutely depends on nucleotide excision repair (NER) and is mechanistically distinct from the replication stress-induced phosphorylation. The treatment of cytosine-β-D-arabinofuranoside strikingly enhances the NER-dependent H2AX phosphorylation and induces the accumulation of replication protein A (RPA) and ATR-interacting protein (ATRIP) at locally UV-damaged subnuclear regions. Consistently, the phosphorylation appears to be mainly mediated by ataxia-telangiectasia mutated and Rad3-related (ATR), although Chk1 (Ser345) is not phosphorylated by the activated ATR. The cellular levels of DNA polymerases δ and ϵ and proliferating cell nuclear antigen are markedly reduced in quiescent cells. We propose a model that perturbed gap-filling synthesis following dual incision in NER generates single-strand DNA gaps and hence initiates H2AX phosphorylation by ATR with the aid of RPA and ATRIP.
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H2AX phosphorylation upon UV radiation is triggered by both NER-dependent and independent pathways.
“Previous studies in this area showed that H2AX phosphorylation upon UV radiation is triggered by both NER-dependent and independent pathways [50;61–63].”
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 [61]
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 [35]
Preventing completion of repair synthesis by genetic or chemical means strongly increases ssDNA accumulation and checkpoint activation.
“Preventing completion of repair synthesis by genetic or chemical means strongly increases accumulation of ssDNA and checkpoint activation, in agreement with a previous report [63].”
From Mind the gap: keeping UV lesions in check (2011) · cites this paper as [63]
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- Authors
- Matsumoto, M., Yaginuma, K., Igarashi, A., Imura, M., Hasegawa, M., Iwabuchi, K., Date, T., Mori, T., Ishizaki, K., Yamashita, K., Inobe, M., & Matsunaga, T.
- Affiliations
- Kanazawa University (JP)
- Kanazawa Medical University (JP)
- Nara Medical University (JP)
- Aichi Cancer Center (JP)
- Corresponding author
- Matsunaga, Tsukasa
- Venue
- Journal of Cell Science
- Publisher
- The Company of Biologists
- Published
- 2007-03-15
- Volume
- 120
- Issue
- 6
- Pages
- 1104-1112
- ISSN
- 0021-9533 (ISSN-L)
- 1477-9137
- OpenAlex
- W2139697903
- Cited by
- 138
- References
- 45
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 4 · 2023: 1 · 2022: 6 · 2021: 6 · 2020: 6 · 2019: 9 · 2018: 5 · 2017: 2 · 2016: 9 · 2015: 5 · 2014: 12 · 2013: 7 · 2012: 13
- FWCI
- 4.74
- Type
- journal article
- Language
- en
- Open access
- Licence
- cc-by-nc-nd
- Locations
- Journal of Cell Science (publishedVersion)
- PubMed (publishedVersion)
- Institutional Repositories DataBase (IRDB) (submittedVersion, cc-by-nc-nd)
- Kanazawa University Repository for Academic Resources (DSpace) (Kanazawa University) (submittedVersion, cc-by-nc-nd)
- 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 › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- H2AX phosphorylation, nucleotide excision repair, UV-induced DNA damage, quiescent cells, ATR, RPA, ATRIP, replication stress, DNA double-strand breaks, DNA polymerase delta, DNA polymerase epsilon, proliferating cell nuclear antigen, cytosine arabinofuranoside, DNA repair signaling
- MeSH terms
- Checkpoint Kinase 1, Cell Cycle, Cells, Cultured, Cytarabine, DNA Damage, DNA Polymerase II, DNA Polymerase III, DNA Repair, DNA-Binding Proteins, DNA, Single-Stranded, Fibroblasts, Histones, Humans, Phosphorylation, Protein Kinases, Ultraviolet Rays, Signal Transduction, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Proliferating Cell Nuclear Antigen, Adaptor Proteins, Signal Transducing, Replication Protein A, Ataxia Telangiectasia Mutated Proteins
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