1. PHF2 regulates homology-directed DNA repair by controlling the resection of DNA double strand breaks
- Author
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Maria Cristina Paz-Cabrera, Pablo Huertas, Cintia Checa-Rodríguez, Juan Ramon Hernandez-Fernaud, Raimundo Freire, Haico van Attikum, Ignacio Alonso-de Vega, Magdalena B. Rother, Wouter W. Wiegant, Veronique A. J. Smits, [Alonso-de Vega,I, Paz-Cabrera,MC, Hernández-Fernaud,JR, Freire,R, Smits,VAJ] Unidad de Investigacion, Hospital Universitario de Canarias, Tenerife, Spain. [Alonso-de Vega,I, Smits,VAJ] Instituto de Tecnologías Biomédicas, Universidad de La Laguna, Tenerife, Spain. [Rother,MB, Wiegant,WW, van Attikum,H] Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. [Checa-Rodríguez,C, Huertas,P] Centro Andaluz de Biologíıa Molecular y Medicina Regenerativa-CABIMER, Sevilla, Spain. [Freire,R, Smits,VAJ] Universidad Fernando Pessoa Canarias, Las Palmas de Gran Canaria, Spain., Ministerio de Ciencia e Innovación [SAF2016-80626-R to V.A.J.S./R.F., SAF2016-74855-P to P.H. and BFU2017-90889-REDT to V.A.J.S./P.H.], co-funded by EU-ERDF, Fundacion Canaria Instituto de Investigación Sanitaria de Canarias (FIISC) [PIFUN16/18 to V.A.J.S.], I.A.V. is supported by a predoctoral fellowship from the Gobierno de Canarias, J.R.H.F. by the Asociacion Española Contra el ˜Cancer, H.v.A. by an ERC Consolidator grant from the European Research Council and a VICI grant from the Netherlands Organisation for Scientific Research. Funding for open access charge: FIISC [PIFUN16/18], Ministerio de Ciencia e Innovacion [SAF2016-80626-R]., Ministerio de Economía y Competitividad (España), Agencia Estatal de Investigación (España), Ministerio de Ciencia, Innovación y Universidades (España), Asociación Española Contra el Cáncer, Fundación Canaria de Investigación Sanitaria, European Research Council, European Commission, Netherlands Organization for Scientific Research, Gobierno de Canarias, Universidad de Sevilla. Departamento de Genética, Universidad de Sevilla. Metabolismo del DNA, Ministerio de Ciencia e Innovación (MICIN). España, and Fundación Canaria Instituto de Investigación Sanitaria de Canarias (FIISC), España
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Genome instability ,AcademicSubjects/SCI00010 ,Phenomena and Processes::Genetic Phenomena::Genetic Processes::Gene Expression Regulation [Medical Subject Headings] ,ADN ,RAD51 ,Genome Integrity, Repair and Replication ,Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings] ,0302 clinical medicine ,Anatomy::Cells::Cells, Cultured::Cell Line [Medical Subject Headings] ,Histonas ,Demethylase activity ,DNA Breaks, Double-Stranded ,Phosphorylation ,Histone Demethylases ,0303 health sciences ,biology ,Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::DNA-Binding Proteins::Homeodomain Proteins [Medical Subject Headings] ,BRCA1 Protein ,Phenomena and Processes::Genetic Phenomena::Genetic Processes::DNA Repair::Recombinational DNA Repair [Medical Subject Headings] ,Double Strand Break Repair ,Chromatin ,Cell biology ,Histone ,Daño del ADN ,030220 oncology & carcinogenesis ,Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Oxidoreductases::Oxidoreductases Acting on CH-NH Group Donors::Oxidoreductases, N-Demethylating::Histone Demethylases [Medical Subject Headings] ,Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Hydrolases::Esterases::Endonucleases::Endodeoxyribonucleases [Medical Subject Headings] ,Fosforilación ,DNA damage ,DNA repair ,Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Nuclear Proteins::BRCA1 Protein [Medical Subject Headings] ,Genomic Instability ,Cell Line ,03 medical and health sciences ,Phenomena and Processes::Genetic Phenomena::Genetic Processes::DNA Damage::DNA Breaks::DNA Breaks, Double-Stranded [Medical Subject Headings] ,Genetics ,Humans ,Proteína BRCA1 ,030304 developmental biology ,Homeodomain Proteins ,Endodeoxyribonucleases ,Recombinational DNA Repair ,DNA ,Phenomena and Processes::Genetic Phenomena::Genomic Instability [Medical Subject Headings] ,Gene Expression Regulation ,Reparación del ADN ,biology.protein ,Anatomy::Cells::Cells, Cultured::Cell Line::Cell Line, Tumor::HeLa Cells [Medical Subject Headings] ,HeLa Cells - Abstract
Post-translational histone modifications and chromatin remodelling play a critical role controlling the integrity of the genome. Here, we identify histone lysine demethylase PHF2 as a novel regulator of the DNA damage response by regulating DNA damage-induced focus formation of 53BP1 and BRCA1, critical factors in the pathway choice for DNA double strand break repair. PHF2 knockdown leads to impaired BRCA1 focus formation and delays the resolution of 53BP1 foci. Moreover, irradiation-induced RPA phosphorylation and focus formation, as well as localization of CtIP, required for DNA end resection, to sites of DNA lesions are affected by depletion of PHF2. These results are indicative of a defective resection of double strand breaks and thereby an impaired homologous recombination upon PHF2 depletion. In accordance with these data, Rad51 focus formation and homology-directed double strand break repair is inhibited in cells depleted for PHF2. Importantly, we demonstrate that PHF2 knockdown decreases CtIP and BRCA1 protein and mRNA levels, an effect that is dependent on the demethylase activity of PHF2. Furthermore, PHF2-depleted cells display genome instability and are mildly sensitive to the inhibition of PARP. Together these results demonstrate that PHF2 promotes DNA repair by homologous recombination by controlling CtIP-dependent resection of double strand breaks., Ministerio de Ciencia e Innovación [SAF2016-80626-R to V.A.J.S./R.F., SAF2016-74855-P to P.H. and BFU2017-90889-REDT to V.A.J.S./P.H.], co-funded by EU-ERDF; Fundación Canaria Instituto de Investigación Sanitaria de Canarias (FIISC) [PIFUN16/18 to V.A.J.S.]; I.A.V. is supported by a predoctoral fellowship from the Gobierno de Canarias; J.R.H.F. by the Asociación Española Contra el Cáncer; H.v.A. by an ERC Consolidator grant from the European Research Council and a VICI grant from the Netherlands Organisation for Scientific Research. Funding for open access charge: FIISC [PIFUN16/18]; Ministerio de Ciencia e Innovación [SAF2016-80626-R].
- Published
- 2020
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