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Rad18 guides polη to replication stalling sites through physical interaction and PCNA monoubiquitination
Open accessAbstract
The DNA replication machinery stalls at damaged sites on templates, but normally restarts by switching to a specialized DNA polymerase(s) that carries out translesion DNA synthesis (TLS). In human cells, DNA polymerase eta (poleta) accumulates at stalling sites as nuclear foci, and is involved in ultraviolet (UV)-induced TLS. Here we show that poleta does not form nuclear foci in RAD18(-/-) cells after UV irradiation. Both Rad18 and Rad6 are required for poleta focus formation. In wild-type cells, UV irradiation induces relocalization of Rad18 in the nucleus, thereby stimulating colocalization with proliferating cell nuclear antigen (PCNA), and Rad18/Rad6-dependent PCNA monoubiquitination. Purified Rad18 and Rad6B monoubiquitinate PCNA in vitro. Rad18 associates with poleta constitutively through domains on their C-terminal regions, and this complex accumulates at the foci after UV irradiation. Furthermore, poleta interacts preferentially with monoubiquitinated PCNA, but poldelta does not. These results suggest that Rad18 is crucial for recruitment of poleta to the damaged site through protein-protein interaction and PCNA monoubiquitination.
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“During translesion synthesis, the PCNA sliding clamp is monoubiquitinated at lysine residue 164 (K164) by the E3 ubiquitin ligase, Rad18 (Watanabe et al., 2004). This modification stimulates recruitment of translesion synthesis polymerases, which bind the PCNA ubiquitin group via their ubiquitin-binding domains (Kannouche et al., 2004; Bienko et al., 2005).”
From USP7 Is a Master Regulator of Genome Stability (2020) · cites this paper as Watanabe et al., 2004
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Metadata
- Authors
- Watanabe, K., Tateishi, S., Kawasuji, M., Tsurimoto, T., Inoue, H., & Yamaizumi, M.
- Affiliations
- Institute of Molecular Embryology and Genetics, Kumamoto University (JP)
- Kumamoto University (JP)
- Kyushu University (JP)
- Saitama University (JP)
- Corresponding author
- Yamaizumi, Masaru
- Venue
- The EMBO Journal
- Published
- 2004-09-09
- Volume
- 23
- Issue
- 19
- Pages
- 3886-3896
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC522788
- OpenAlex
- W2020136215
- Cited by
- 538
- References
- 34
- Citations by year
- 2026: 5 · 2025: 15 · 2024: 22 · 2023: 15 · 2022: 16 · 2021: 18 · 2020: 18 · 2019: 14 · 2018: 27 · 2017: 18 · 2016: 24 · 2015: 21 · 2014: 19 · 2013: 23 · 2012: 37
- FWCI
- 11.96
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- The EMBO Journal (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Princess Takamatsu Cancer Research Fund
- 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
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- RAD18, DNA polymerase eta, PCNA monoubiquitination, DNA replication stalling, translesion synthesis, UV-induced DNA damage, replication restart, protein-protein interaction, PCNA, DNA polymerase delta, nuclear foci, DNA damage response
- MeSH terms
- Y-Family DNA Polymerases, Animals, Cell Nucleus, DNA Polymerase III, DNA-Directed DNA Polymerase, DNA Replication, DNA-Binding Proteins, Female, Cricetinae, Humans, Male, Mice, Inbred C57BL, Ultraviolet Rays, CHO Cells, Proliferating Cell Nuclear Antigen, Ubiquitin, Ubiquitin-Conjugating Enzymes, Ubiquitin-Protein Ligases, Mice
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