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

Julianne Kitevski-LeBlanc; Amélie Fradet-Turcotte; Predrag Kukic; Marcus D Wilson; Guillem Portella; Tairan Yuwen; Stephanie Panier; Shili Duan; Marella D Canny; Hugo van Ingen; Cheryl H Arrowsmith; John L Rubinstein; Michele Vendruscolo; Daniel Durocher; Lewis E Kay

doi:10.7554/eLife.23872

eLife (Apr 2017)

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

Julianne Kitevski-LeBlanc,
Amélie Fradet-Turcotte,
Predrag Kukic,
Marcus D Wilson,
Guillem Portella,
Tairan Yuwen,
Stephanie Panier,
Shili Duan,
Marella D Canny,
Hugo van Ingen,
Cheryl H Arrowsmith,
John L Rubinstein,
Michele Vendruscolo,
Daniel Durocher,
Lewis E Kay

Affiliations

Julianne Kitevski-LeBlanc: ORCiD; Department of Molecular Genetics, University of Toronto, Toronto, Canada; Department of Biochemistry, University of Toronto, Toronto, Canada; Department of Chemistry, University of Toronto, Toronto, Canada
Amélie Fradet-Turcotte: ORCiD; The Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada; Laval University Cancer Research Center, Oncology Axis – Centre Hospitalier Universitaire de Québec Research Center – Université Laval, Hôtel-Dieu de Québec, Québec City, Canada
Predrag Kukic: Department of Chemistry, University of Cambridge, Cambridge, United Kingdom
Marcus D Wilson: ORCiD; The Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada
Guillem Portella: Department of Chemistry, University of Cambridge, Cambridge, United Kingdom
Tairan Yuwen: ORCiD; Department of Molecular Genetics, University of Toronto, Toronto, Canada; Department of Biochemistry, University of Toronto, Toronto, Canada; Department of Chemistry, University of Toronto, Toronto, Canada
Stephanie Panier: Department of Molecular Genetics, University of Toronto, Toronto, Canada; The Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada
Shili Duan: Structural Genomics Consortium, University of Toronto, Toronto, Canada; Princess Margret Cancer Centre, Toronto, Canada
Marella D Canny: Department of Medical Biophysics, University of Toronto, Toronto, Canada
Hugo van Ingen: Macromolecular Biochemistry, Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands
Cheryl H Arrowsmith: Structural Genomics Consortium, University of Toronto, Toronto, Canada; Princess Margret Cancer Centre, Toronto, Canada; Department of Medical Biophysics, University of Toronto, Toronto, Canada
John L Rubinstein: ORCiD; Department of Molecular Genetics, University of Toronto, Toronto, Canada; Department of Medical Biophysics, University of Toronto, Toronto, Canada; Molecular Structure and Function Program, The Hospital for Sick Children Research Institute, Toronto, Canada
Michele Vendruscolo: ORCiD; Department of Chemistry, University of Cambridge, Cambridge, United Kingdom
Daniel Durocher: ORCiD; Department of Molecular Genetics, University of Toronto, Toronto, Canada; The Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada
Lewis E Kay: ORCiD; Department of Molecular Genetics, University of Toronto, Toronto, Canada; Department of Biochemistry, University of Toronto, Toronto, Canada; Department of Chemistry, University of Toronto, Toronto, Canada; Molecular Structure and Function Program, The Hospital for Sick Children Research Institute, Toronto, Canada

DOI: https://doi.org/10.7554/eLife.23872
Journal volume & issue: Vol. 6

Abstract

Read online

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.

Published in eLife

ISSN: 2050-084X (Online)
Publisher: eLife Sciences Publications Ltd
Country of publisher: United Kingdom
LCC subjects: Medicine; Science: Biology (General)
Website: https://elifesciences.org

About the journal

Abstract

Keywords