title: SETD2-dependent histone H3K36 trimethylation is required for homologous recombination repair and genome stability.
abstract: Modulating chromatin through histone methylation orchestrates numerous cellular processes. SETD2-dependent trimethylation of histone H3K36 is associated with active transcription. Here, we define a role for H3K36 trimethylation in homologous recombination (HR) repair in human cells. We find that depleting SETD2 generates a mutation signature resembling RAD51 depletion at I-SceI-induced DNA double-strand break (DSB) sites, with significantly increased deletions arising through microhomology-mediated end-joining. We establish a presynaptic role for SETD2 methyltransferase in HR, where it facilitates the recruitment of C-terminal binding protein interacting protein (CtIP) and promotes DSB resection, allowing Replication Protein A (RPA) and RAD51 binding to DNA damage sites. Furthermore, reducing H3K36me3 levels by overexpressing KDM4A/JMJD2A, an oncogene and H3K36me3/2 demethylase, or an H3.3K36M transgene also reduces HR repair events. We propose that error-free HR repair within H3K36me3-decorated transcriptionally active genomic regions promotes cell homeostasis. Moreover, these findings provide insights as to why oncogenic mutations cluster within the H3K36me3 axis.
authors.affiliation: CRUK MRC Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK.
authors.forename: Sophia X
authors.initials: SX
authors.lastname: Pfister
authors.affiliation: CRUK MRC Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK.
authors.forename: Sara
authors.initials: S
authors.lastname: Ahrabi
authors.affiliation: Laboratory of Cancer Biology, Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK.
authors.forename: Lykourgos-Panagiotis
authors.initials: LP
authors.lastname: Zalmas
authors.affiliation: CRUK MRC Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK.
authors.forename: Sovan
authors.initials: S
authors.lastname: Sarkar
authors.affiliation: Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération, Université de Toulouse, Université Paul Sabatier, 31062 Toulouse, France; CNRS, Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération, 31062 Toulouse, France.
authors.forename: François
authors.initials: F
authors.lastname: Aymard
authors.affiliation: School of Life Sciences, University of Lincoln, Lincoln LN6 7TS, UK.
authors.forename: Csanád Z
authors.initials: CZ
authors.lastname: Bachrati
authors.affiliation: Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 21 Stockholm, Sweden.
authors.forename: Thomas
authors.initials: T
authors.lastname: Helleday
authors.affiliation: Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération, Université de Toulouse, Université Paul Sabatier, 31062 Toulouse, France; CNRS, Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération, 31062 Toulouse, France.
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Legube
authors.affiliation: Laboratory of Cancer Biology, Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK.
authors.forename: Nicholas B
authors.initials: NB
authors.lastname: La Thangue
authors.affiliation: Gene Targeting Group, Centre for Haematology, Imperial College Faculty of Medicine, London W12 0NN, UK.
authors.forename: Andrew C G
authors.initials: AC
authors.lastname: Porter
authors.affiliation: CRUK MRC Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK. Electronic address: timothy.humphrey@oncology.ox.ac.uk.
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Humphrey
mesh_terms: D004260:DNA Repair; D042822:Genomic Instability; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics* / Q000378:metabolism; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D059765:Homologous Recombination; D006801:Humans; D008745:Methylation; D011485:Protein Binding; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D059767:Recombinational DNA Repair; D014162:Transfection
chemical_list: D006657:Histones; D011495:Histone-Lysine N-Methyltransferase; C521636:SETD2 protein, human; C495153:RAD51 protein, human; D051135:Rad51 Recombinase