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Cell Rep. 2015 Jun 9;11(9):1486-500. doi: 10.1016/j.celrep.2015.04.053. Epub 2015 May 21.

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

Author information

1
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.
2
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.
3
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.
4
Department of Cell Biology, Harvard University Medical School, Boston, MA 02115, USA.
5
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.
6
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.

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.

PMID:
26004182
PMCID:
PMC4464939
DOI:
10.1016/j.celrep.2015.04.053
[Indexed for MEDLINE]
Free PMC Article

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