title: Fine-tuning of DNA damage-dependent ubiquitination by OTUB2 supports the DNA repair pathway choice.
abstract: DNA double-strand breaks (DSBs) are deleterious lesions that lead to genetic mutations and cell death. Protein ubiquitination mediated by the E3 ubiquitin ligase RNF8 within the regions surrounding DSBs recruits DNA DSB response (DDR) factors and induces chromatin remodeling, which supports cell survival after DNA damage. Nevertheless, the impact of RNF8-mediated ubiquitination on DNA repair remains to be elucidated. Here, we report that depletion of the deubiquitinating enzyme OTUB2 enhances RNF8-mediated ubiquitination in an early phase of the DDR and promotes faster DSB repair but suppresses homologous recombination. The rapid ubiquitination results in accelerated accumulation of 53BP1 and RAP80 at DSBs, which in turn protects DSB ends from resection in OTUB2-depleted cells. Mechanistically, OTUB2 suppresses RNF8-mediated L3MBTL1 ubiquitination and Lys 63-linked ubiquitin chain formation in a deubiquitinating activity-dependent manner. Thus, OTUB2 fine-tunes the speed of DSB-induced ubiquitination so that the appropriate DNA repair pathway is chosen.
authors.affiliation: Department of Bioregulation and Cellular Response, Graduate School of Medicine, Osaka University, 2-2Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Kiyoko
authors.initials: K
authors.lastname: Kato
authors.affiliation: Department of Bioregulation and Cellular Response, Graduate School of Medicine, Osaka University, 2-2Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Nakajima
authors.affiliation: Division of Dynamic Proteome in Aging and Cancer, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryomachi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Ui
authors.affiliation: Center for Medical Research and Education, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Muto-Terao
authors.affiliation: Division of Genome Biology, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, 104-0045, Tokyo, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Ogiwara
authors.affiliation: Department of Bioregulation and Cellular Response, Graduate School of Medicine, Osaka University, 2-2Yamadaoka, Suita, Osaka, 565-0871, Japan. Electronic address: snakada@bcr.med.osaka-u.ac.jp.
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Nakada
mesh_terms: D002352:Carrier Proteins / Q000378:metabolism; D016923:Cell Death; D045744:Cell Line, Tumor; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000737:chemistry; D015723:Gene Library; D020868:Gene Silencing; D006367:HeLa Cells; D056488:Histone Chaperones; D006657:Histones / Q000737:chemistry; D059765:Homologous Recombination; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008239:Lysine / Q000737:chemistry; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D011995:Recombination, Genetic; D013869:Thiolester Hydrolases / Q000737:chemistry*; D000071857:Tumor Suppressor p53-Binding Protein 1; D025801:Ubiquitin / Q000737:chemistry; D044767:Ubiquitin-Protein Ligases
chemical_list: D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D056488:Histone Chaperones; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; C486521:RNF8 protein, human; C498301:TP53BP1 protein, human; D000071857:Tumor Suppressor p53-Binding Protein 1; C466234:UIMC1 protein, human; D025801:Ubiquitin; D044767:Ubiquitin-Protein Ligases; C496399:OTUB2 protein, human; D013869:Thiolester Hydrolases; D008239:Lysine