title: A Systematic Analysis of Factors Localized to Damaged Chromatin Reveals PARP-Dependent Recruitment of Transcription Factors.
abstract: Localization to sites of DNA damage is a hallmark of DNA damage response (DDR) proteins. To identify DDR factors, we screened epitope-tagged proteins for localization to sites of chromatin damaged by UV laser microirradiation and found >120 proteins that localize to damaged chromatin. These include the BAF tumor suppressor complex and the amyotrophic lateral sclerosis (ALS) candidate protein TAF15. TAF15 contains multiple domains that bind damaged chromatin in a poly-(ADP-ribose) polymerase (PARP)-dependent manner, suggesting a possible role as glue that tethers multiple PAR chains together. Many positives were transcription factors; > 70% of randomly tested transcription factors localized to sites of DNA damage, and of these, ∼90% were PARP dependent for localization. Mutational analyses showed that localization to damaged chromatin is DNA-binding-domain dependent. By examining Hoechst staining patterns at damage sites, we see evidence of chromatin decompaction that is PARP dependent. We propose that PARP-regulated chromatin remodeling at sites of damage allows transient accessibility of DNA-binding proteins.
authors.affiliation: Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute; Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Lior
authors.initials: L
authors.lastname: Izhar
authors.affiliation: Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute; Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Britt
authors.initials: B
authors.lastname: Adamson
authors.affiliation: Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute; Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ciccia
authors.affiliation: Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute; Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Jedd
authors.initials: J
authors.lastname: Lewis
authors.affiliation: Department of Cell Biology, Harvard University Medical School, Boston, MA 02115, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Pontano-Vaites
authors.affiliation: Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute; Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Yumei
authors.initials: Y
authors.lastname: Leng
authors.affiliation: Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute; Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Anthony C
authors.initials: AC
authors.lastname: Liang
authors.affiliation: Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Department of Molecular and Human Genetics, and Department of Pediatrics, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Westbrook
authors.affiliation: Department of Cell Biology, Harvard University Medical School, Boston, MA 02115, USA.
authors.forename: J Wade
authors.initials: JW
authors.lastname: Harper
authors.affiliation: Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute; Division of Genetics, Brigham and Women's Hospital, Boston, MA 02115, USA. Electronic address: selledge@genetics.med.harvard.edu.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Elledge
mesh_terms: D002843:Chromatin / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly / Q000502:physiology*; D004249:DNA Damage / Q000502:physiology*; D004260:DNA Repair / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D057809:HEK293 Cells; D006801:Humans; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D002843:Chromatin; D014157:Transcription Factors; D011065:Poly(ADP-ribose) Polymerases