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Elife. 2017 Apr 13;6. pii: e23872. doi: 10.7554/eLife.23872.

The RNF168 paralog RNF169 defines a new class of ubiquitylated histone reader involved in the response to DNA damage.

Author information

1
Department of Molecular Genetics, University of Toronto, Toronto, Canada.
2
Department of Biochemistry, University of Toronto, Toronto, Canada.
3
Department of Chemistry, University of Toronto, Toronto, Canada.
4
The Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada.
5
Laval University Cancer Research Center, Oncology Axis - Centre Hospitalier Universitaire de Québec Research Center - Université Laval, Hôtel-Dieu de Québec, Québec City, Canada.
6
Department of Chemistry, University of Cambridge, Cambridge, United Kingdom.
7
Structural Genomics Consortium, University of Toronto, Toronto, Canada.
8
Princess Margret Cancer Centre, Toronto, Canada.
9
Department of Medical Biophysics, University of Toronto, Toronto, Canada.
10
Macromolecular Biochemistry, Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands.
11
Molecular Structure and Function Program, The Hospital for Sick Children Research Institute, Toronto, Canada.

Abstract

Site-specific histone ubiquitylation plays a central role in orchestrating the response to DNA double-strand breaks (DSBs). DSBs elicit a cascade of events controlled by the ubiquitin ligase RNF168, which promotes the accumulation of repair factors such as 53BP1 and BRCA1 on the chromatin flanking the break site. RNF168 also promotes its own accumulation, and that of its paralog RNF169, but how they recognize ubiquitylated chromatin is unknown. Using methyl-TROSY solution NMR spectroscopy and molecular dynamics simulations, we present an atomic resolution model of human RNF169 binding to a ubiquitylated nucleosome, and validate it by electron cryomicroscopy. We establish that RNF169 binds to ubiquitylated H2A-Lys13/Lys15 in a manner that involves its canonical ubiquitin-binding helix and a pair of arginine-rich motifs that interact with the nucleosome acidic patch. This three-pronged interaction mechanism is distinct from that by which 53BP1 binds to ubiquitylated H2A-Lys15 highlighting the diversity in site-specific recognition of ubiquitylated nucleosomes.

KEYWORDS:

D. melanogaster; DNA damage; biophysics; human; structural biology; ubiquitin-based signalling; ubiquitylated-histone reader

PMID:
28406400
PMCID:
PMC5426901
DOI:
10.7554/eLife.23872
[Indexed for MEDLINE]
Free PMC Article

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