title: RNA m6A methylation regulates the ultraviolet-induced DNA damage response.
abstract: Cell proliferation and survival require the faithful maintenance and propagation of genetic information, which are threatened by the ubiquitous sources of DNA damage present intracellularly and in the external environment. A system of DNA repair, called the DNA damage response, detects and repairs damaged DNA and prevents cell division until the repair is complete. Here we report that methylation at the 6 position of adenosine (m6A) in RNA is rapidly (within 2 min) and transiently induced at DNA damage sites in response to ultraviolet irradiation. This modification occurs on numerous poly(A)+transcripts and is regulated by the methyltransferase METTL3 (methyltransferase-like 3) and the demethylase FTO (fat mass and obesity-associated protein). In the absence of METTL3 catalytic activity, cells showed delayed repair of ultraviolet-induced cyclobutane pyrimidine adducts and elevated sensitivity to ultraviolet, demonstrating the importance of m6A in the ultraviolet-responsive DNA damage response. Multiple DNA polymerases are involved in the ultraviolet response, some of which resynthesize DNA after the lesion has been excised by the nucleotide excision repair pathway, while others participate in trans-lesion synthesis to allow replication past damaged lesions in S phase. DNA polymerase κ (Pol κ), which has been implicated in both nucleotide excision repair and trans-lesion synthesis, required the catalytic activity of METTL3 for immediate localization to ultraviolet-induced DNA damage sites. Importantly, Pol κ overexpression qualitatively suppressed the cyclobutane pyrimidine removal defect associated with METTL3 loss. Thus, we have uncovered a novel function for RNA m6A modification in the ultraviolet-induced DNA damage response, and our findings collectively support a model in which m6A RNA serves as a beacon for the selective, rapid recruitment of Pol κ to damage sites to facilitate repair and cell survival.
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xiang
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Benoit
authors.initials: B
authors.lastname: Laurent
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Chih-Hung
authors.initials: CH
authors.lastname: Hsu
authors.affiliation: Department of Chemistry, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.|Institute for Biophysical Dynamics, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.|Howard Hughes Medical Institute, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.
authors.forename: Sigrid
authors.initials: S
authors.lastname: Nachtergaele
authors.affiliation: Department of Chemistry, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.|Institute for Biophysical Dynamics, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.|Howard Hughes Medical Institute, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.
authors.forename: Zhike
authors.initials: Z
authors.lastname: Lu
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Wanqiang
authors.initials: W
authors.lastname: Sheng
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Chuanyun
authors.initials: C
authors.lastname: Xu
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, Massachusetts 02109, USA.|Department of Pathology, Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, Massachusetts 02109, USA.
authors.forename: Jian
authors.initials: J
authors.lastname: Ouyang
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Siqing
authors.initials: S
authors.lastname: Wang
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Dominic
authors.initials: D
authors.lastname: Ling
authors.affiliation: Department of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung City 202, Taiwan.
authors.forename: Pang-Hung
authors.initials: PH
authors.lastname: Hsu
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, Massachusetts 02109, USA.|Department of Pathology, Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, Massachusetts 02109, USA.
authors.forename: Lee
authors.initials: L
authors.lastname: Zou
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Ashwini
authors.initials: A
authors.lastname: Jambhekar
authors.affiliation: Department of Chemistry, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.|Institute for Biophysical Dynamics, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.|Howard Hughes Medical Institute, University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.
authors.forename: Chuan
authors.initials: C
authors.lastname: He
authors.affiliation: Division of Newborn Medicine and Epigenetics Program, Department of Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Shi
mesh_terms: D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO / Q000378:metabolism; D000818:Animals; D055162:Biocatalysis / Q000528:radiation effects; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D004249:DNA Damage / Q000528:radiation effects*; D004260:DNA Repair / Q000528:radiation effects; D004261:DNA Replication / Q000528:radiation effects; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D006801:Humans; D008745:Methylation / Q000528:radiation effects; D008780:Methyltransferases / Q000172:deficiency / Q000378:metabolism; D051379:Mice; D011061:Poly A / Q000378:metabolism; D012313:RNA / Q000737:chemistry* / Q000378:metabolism* / Q000528:radiation effects; D016196:S Phase / Q000528:radiation effects; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
chemical_list: D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO; D004259:DNA-Directed DNA Polymerase; D008780:Methyltransferases; D011061:Poly A; D012313:RNA; C526492:FTO protein, human; C468275:METTL3 protein, human; C404302:POLK protein, human; D000099270:Y-Family DNA Polymerases