A PALB2-interacting domain in RNF168 couples homologous recombination to DNA break-induced chromatin ubiquitylation

Martijn S Luijsterburg; Dimitris Typas; Marie-Christine Caron; Wouter W Wiegant; Diana van den Heuvel; Rick A Boonen; Anthony M Couturier; Leon H Mullenders; Jean-Yves Masson; Haico van Attikum

doi:10.7554/eLife.20922

A PALB2-interacting domain in RNF168 couples homologous recombination to DNA break-induced chromatin ubiquitylation

Elife. 2017 Feb 27:6:e20922. doi: 10.7554/eLife.20922.

Authors

Affiliations

¹ Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
² Genome Stability Laboratory, CHU de Québec Research Center, HDQ Pavilion, Oncology Axis, McMahon, Québec City, Canada.
³ Department of Molecular Biology, Medical Biochemistry and Pathology, Laval University Cancer Research Center, Québec City, Canada.

Abstract

DNA double-strand breaks (DSB) elicit a ubiquitylation cascade that controls DNA repair pathway choice. This cascade involves the ubiquitylation of histone H2A by the RNF168 ligase and the subsequent recruitment of RIF1, which suppresses homologous recombination (HR) in G1 cells. The RIF1-dependent suppression is relieved in S/G2 cells, allowing PALB2-driven HR to occur. With the inhibitory impact of RIF1 relieved, it remains unclear how RNF168-induced ubiquitylation influences HR. Here, we uncover that RNF168 links the HR machinery to H2A ubiquitylation in S/G2 cells. We show that PALB2 indirectly recognizes histone ubiquitylation by physically associating with ubiquitin-bound RNF168. This direct interaction is mediated by the newly identified PALB2-interacting domain (PID) in RNF168 and the WD40 domain in PALB2, and drives DNA repair by facilitating the assembly of PALB2-containing HR complexes at DSBs. Our findings demonstrate that RNF168 couples PALB2-dependent HR to H2A ubiquitylation to promote DNA repair and preserve genome integrity.

Keywords: DNA repair; PALB2; RNF168; biochemistry; cell biology; chromatin; homologous recombination; human; ubiquitylation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Cycle / genetics
Cell Cycle / radiation effects
Cell Line, Transformed
Cell Line, Tumor
DNA / genetics
DNA / metabolism*
DNA Breaks, Double-Stranded / radiation effects
Fanconi Anemia Complementation Group N Protein / genetics
Fanconi Anemia Complementation Group N Protein / metabolism*
Fibroblasts / cytology
Fibroblasts / metabolism*
Fibroblasts / radiation effects
HEK293 Cells
Histones / genetics
Histones / metabolism*
Humans
Lasers, Excimer
Mice
Mouse Embryonic Stem Cells / cytology
Mouse Embryonic Stem Cells / metabolism
Mouse Embryonic Stem Cells / radiation effects
Osteoblasts / cytology
Osteoblasts / metabolism
Osteoblasts / radiation effects
Protein Binding
Protein Interaction Domains and Motifs
Recombinational DNA Repair*
Telomere-Binding Proteins / genetics
Telomere-Binding Proteins / metabolism
Ubiquitin / genetics
Ubiquitin / metabolism
Ubiquitin-Protein Ligases / genetics
Ubiquitin-Protein Ligases / metabolism*
Ubiquitination
X-Rays

Substances

Fanconi Anemia Complementation Group N Protein
Histones
PALB2 protein, human
Rif1 protein, human
Telomere-Binding Proteins
Ubiquitin
DNA
RNF168 protein, human
Ubiquitin-Protein Ligases

Grants and funding

CIHR/Canada