Cited by 538DNA Repair MechanismsUbiquitin and proteasome pathwaysDNA and Nucleic Acid Chemistry

Rad18 guides polη to replication stalling sites through physical interaction and PCNA monoubiquitination

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Watanabe, Kenji, Tateishi, Satoshi, Kawasuji, Michio, Tsurimoto, Toshiki, Inoue, Hirokazu, Yamaizumi, Masaru·2004·The EMBO Journal

Abstract

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, PCNA is monoubiquitinated at K164 by Rad18, stimulating recruitment of TLS polymerases.

    “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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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
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
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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