A Systematic Analysis of Factors Localized to Damaged Chromatin Reveals PARP-Dependent Recruitment of Transcription Factors.

TitleA Systematic Analysis of Factors Localized to Damaged Chromatin Reveals PARP-Dependent Recruitment of Transcription Factors.
Publication TypeJournal Article
Year of Publication2015
AuthorsIzhar, L, Adamson, B, Ciccia, A, Lewis, J, Pontano-Vaites, L, Leng, Y, Liang, AC, Westbrook, TF, J Harper, W, Elledge, SJ
JournalCell Rep
Volume11
Issue9
Pagination1486-500
Date Published2015 Jun 09
ISSN2211-1247
KeywordsChromatin, Chromatin Assembly and Disassembly, DNA Damage, DNA Repair, Fluorescent Antibody Technique, HEK293 Cells, Humans, Poly(ADP-ribose) Polymerases, 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.

DOI10.1016/j.celrep.2015.04.053
Alternate JournalCell Rep
PubMed ID26004182
PubMed Central IDPMC4464939
Grant ListGM44664 / GM / NIGMS NIH HHS / United States
AG011085 / AG / NIA NIH HHS / United States
R01 GM044664 / GM / NIGMS NIH HHS / United States
R01 CA178039 / CA / NCI NIH HHS / United States
R01 AG011085 / AG / NIA NIH HHS / United States
/ / Howard Hughes Medical Institute / United States
R37 GM044664 / GM / NIGMS NIH HHS / United States
1R01CA178039 / CA / NCI NIH HHS / United States