title: Mammalian RAD52 Functions in Break-Induced Replication Repair of Collapsed DNA Replication Forks.
abstract: Human cancers are characterized by the presence of oncogene-induced DNA replication stress (DRS), making them dependent on repair pathways such as break-induced replication (BIR) for damaged DNA replication forks. To better understand BIR, we performed a targeted siRNA screen for genes whose depletion inhibited G1 to S phase progression when oncogenic cyclin E was overexpressed. RAD52, a gene dispensable for normal development in mice, was among the top hits. In cells in which fork collapse was induced by oncogenes or chemicals, the Rad52 protein localized to DRS foci. Depletion of Rad52 by siRNA or knockout of the gene by CRISPR/Cas9 compromised restart of collapsed forks and led to DNA damage in cells experiencing DRS. Furthermore, in cancer-prone, heterozygous APC mutant mice, homozygous deletion of the Rad52 gene suppressed tumor growth and prolonged lifespan. We therefore propose that mammalian RAD52 facilitates repair of collapsed DNA replication forks in cancer cells.
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland.
authors.forename: Sotirios K
authors.initials: SK
authors.lastname: Sotiriou
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland.
authors.forename: Irene
authors.initials: I
authors.lastname: Kamileri
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Lugli
authors.affiliation: Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 75 Mikras Asias Street, 11527 Athens, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Evangelou
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland.
authors.forename: Caterina
authors.initials: C
authors.lastname: Da-Ré
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland.
authors.forename: Florian
authors.initials: F
authors.lastname: Huber
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland.
authors.forename: Laura
authors.initials: L
authors.lastname: Padayachy
authors.affiliation: School of Pharmaceutical Sciences, Department of Pharmaceutical Biochemistry, CMU, University of Geneva and University of Lausanne, Rue Michel-Servet 1, 1211 Geneva, Switzerland.
authors.forename: Sebastien
authors.initials: S
authors.lastname: Tardy
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland; School of Pharmaceutical Sciences, Department of Pharmaceutical Biochemistry, CMU, University of Geneva and University of Lausanne, Rue Michel-Servet 1, 1211 Geneva, Switzerland.
authors.forename: Noemie L
authors.initials: NL
authors.lastname: Nicati
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland.
authors.forename: Samia
authors.initials: S
authors.lastname: Barriot
authors.affiliation: Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.
authors.forename: Fena
authors.initials: F
authors.lastname: Ochs
authors.affiliation: Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.
authors.forename: Claudia
authors.initials: C
authors.lastname: Lukas
authors.affiliation: Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.
authors.forename: Jiri
authors.initials: J
authors.lastname: Lukas
authors.affiliation: Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 75 Mikras Asias Street, 11527 Athens, Greece; Faculty Institute for Cancer Sciences, Manchester Academic Health Sciences Centre, University of Manchester, Manchester M13 9PL, UK; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece.
authors.forename: Vassilis G
authors.initials: VG
authors.lastname: Gorgoulis
authors.affiliation: School of Pharmaceutical Sciences, Department of Pharmaceutical Biochemistry, CMU, University of Geneva and University of Lausanne, Rue Michel-Servet 1, 1211 Geneva, Switzerland.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Scapozza
authors.affiliation: Department of Molecular Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211 Geneva, Switzerland. Electronic address: thanos.halazonetis@unige.ch.
authors.forename: Thanos D
authors.initials: TD
authors.lastname: Halazonetis
mesh_terms: D025601:Adenomatous Polyposis Coli Protein / Q000172:deficiency / Q000235:genetics*; D000818:Animals; D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D019927:Cyclin E / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D016193:G1 Phase; D015870:Gene Expression; D042822:Genomic Instability; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D015739:Nocodazole / Q000494:pharmacology; D012516:Osteosarcoma / Q000235:genetics* / Q000378:metabolism / Q000401:mortality / Q000473:pathology; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D051721:Rad52 DNA Repair and Recombination Protein / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D059767:Recombinational DNA Repair; D016196:S Phase; D013312:Stress, Physiological; D016019:Survival Analysis
chemical_list: D025601:Adenomatous Polyposis Coli Protein; D019927:Cyclin E; D034741:RNA, Small Interfering; D051721:Rad52 DNA Repair and Recombination Protein; C497196:Rad52 protein, mouse; C000613192:adenomatous polyposis coli protein, mouse; D004247:DNA; D015739:Nocodazole
keywords: DNA recombination; DNA replication stress; RAD52; break-induced replication; cancer