title: The role of HERC2 and RNF8 ubiquitin E3 ligases in the promotion of translesion DNA synthesis in the chicken DT40 cell line.
abstract: The replicative DNA polymerases are generally blocked by template DNA damage. The resulting replication arrest can be released by one of two post-replication repair (PRR) pathways, translesion DNA synthesis (TLS) and template switching by homologous recombination (HR). The HERC2 ubiquitin ligase plays a role in homologous recombination by facilitating the assembly of the Ubc13 ubiquitin-conjugating enzyme with the RNF8 ubiquitin ligase. To explore the role of HERC2 and RNF8 in PRR, we examined immunoglobulin diversification in chicken DT40 cells deficient in HERC2 and RNF8. Unexpectedly, the HERC2(-/-) and RNF8(-/-) cells and HERC2(-/-)/RNF8(-/-) double mutant cells exhibit a significant reduction in the rate of immunoglobulin (Ig) hypermutation, compared to wild-type cells. Further, the HERC2(-/-) and RNF8(-/-) mutants exhibit defective maintenance of replication fork progression immediately after exposure to UV while retaining proficient post-replicative gap filling. These mutants are both proficient in mono-ubiquitination of PCNA. Taken together, these results suggest that HERC2 and RNF8 promote TLS past abasic sites and UV-lesions at or very close to stalled replication forks.
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan.
authors.lastname: Mohiuddin
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Islam Shamima
authors.initials: IS
authors.lastname: Keka
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Guilbaud
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK.
authors.forename: Julian
authors.initials: J
authors.lastname: Sale
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan; Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
authors.forename: Takeo
authors.initials: T
authors.lastname: Narita
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan; Faculty of Medicine, Seuz Canal University, circular road Ez-Eldeen, Ismailia 41522, Egypt.
authors.forename: H Ismail
authors.initials: HI
authors.lastname: Abdel-Aziz
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Saki
authors.initials: S
authors.lastname: Ogawa
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sasanuma
authors.affiliation: University College Groningen, University of Groningen, 9718 BG Groningen, Hoendiepskade 23-24, The Netherlands.
authors.forename: Roland
authors.initials: R
authors.lastname: Chiu
authors.affiliation: Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
authors.forename: Vibe H
authors.initials: VH
authors.lastname: Oestergaard
authors.affiliation: Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
authors.forename: Michael
authors.initials: M
authors.lastname: Lisby
authors.affiliation: Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan. Electronic address: stakeda@rg.med.kyoto-u.ac.jp.
authors.forename: Shunichi
authors.initials: S
authors.lastname: Takeda
mesh_terms: D000818:Animals; D002645:Chickens; D004249:DNA Damage; D004261:DNA Replication; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination; D014466:Ultraviolet Rays
chemical_list: D020662:Guanine Nucleotide Exchange Factors; D044767:Ubiquitin-Protein Ligases
keywords: HERC2; Post-replication repair; RNF8; Translesion DNA synthesis; Ubiquitination