MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair

Inge de Krijger; Bastian Föhr; Santiago Hernández Pérez; Estelle Vincendeau; Judit Serrat; Alexander Marc Thouin; Vivek Susvirkar; Chloé Lescale; Inés Paniagua; Liesbeth Hoekman; Simranjeet Kaur; Maarten Altelaar; Ludovic Deriano; Alex C Faesen; Jacqueline J L Jacobs

doi:10.1038/s41467-021-25724-y

MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair

Nat Commun. 2021 Sep 14;12(1):5421. doi: 10.1038/s41467-021-25724-y.

Authors

Inge de Krijger¹, Bastian Föhr^#², Santiago Hernández Pérez^#¹, Estelle Vincendeau^#^{3

4}, Judit Serrat^#¹, Alexander Marc Thouin^#¹, Vivek Susvirkar², Chloé Lescale³, Inés Paniagua¹, Liesbeth Hoekman⁵, Simranjeet Kaur², Maarten Altelaar^{5

6}, Ludovic Deriano³, Alex C Faesen², Jacqueline J L Jacobs⁷

Affiliations

¹ Division of Oncogenomics, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
² Laboratory of Signal Dynamics, Max-Planck Institute for Biophysical Chemistry, Göttingen, Germany.
³ Genome Integrity, Immunity and Cancer Unit, Equipe Labellisée Ligue Contre Le Cancer, INSERM U1223, Institut Pasteur, Paris, France.
⁴ Université de Paris, Sorbonne Paris Cité, Paris, France.
⁵ Proteomics Facility, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
⁶ Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Utrecht, The Netherlands.
⁷ Division of Oncogenomics, The Netherlands Cancer Institute, Amsterdam, The Netherlands. j.jacobs@nki.nl.

^# Contributed equally.

Abstract

MAD2L2 (REV7) plays an important role in DNA double-strand break repair. As a member of the shieldin complex, consisting of MAD2L2, SHLD1, SHLD2 and SHLD3, it controls DNA repair pathway choice by counteracting DNA end-resection. Here we investigated the requirements for shieldin complex assembly and activity. Besides a dimerization-surface, HORMA-domain protein MAD2L2 has the extraordinary ability to wrap its C-terminus around SHLD3, likely creating a very stable complex. We show that appropriate function of MAD2L2 within shieldin requires its dimerization, mediated by SHLD2 and accelerating MAD2L2-SHLD3 interaction. Dimerization-defective MAD2L2 impairs shieldin assembly and fails to promote NHEJ. Moreover, MAD2L2 dimerization, along with the presence of SHLD3, allows shieldin to interact with the TRIP13 ATPase, known to drive topological switches in HORMA-domain proteins. We find that appropriate levels of TRIP13 are important for proper shieldin (dis)assembly and activity in DNA repair. Together our data provide important insights in the dependencies for shieldin activity.

Publication types

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

MeSH terms

ATPases Associated with Diverse Cellular Activities / chemistry
ATPases Associated with Diverse Cellular Activities / genetics*
ATPases Associated with Diverse Cellular Activities / metabolism
Animals
Binding Sites
Cell Cycle Proteins / chemistry
Cell Cycle Proteins / genetics*
Cell Cycle Proteins / metabolism
Cell Line
Cell Line, Tumor
Cisplatin / pharmacology
DNA / chemistry
DNA / genetics*
DNA / metabolism
DNA Breaks, Double-Stranded
DNA Repair*
DNA-Binding Proteins / chemistry
DNA-Binding Proteins / genetics*
DNA-Binding Proteins / metabolism
Fibroblasts / cytology
Fibroblasts / drug effects
Fibroblasts / metabolism
Gene Expression
HEK293 Cells
HeLa Cells
Humans
Mad2 Proteins / chemistry
Mad2 Proteins / genetics*
Mad2 Proteins / metabolism
Mice
Phthalazines / pharmacology
Piperazines / pharmacology
Protein Binding
Protein Interaction Domains and Motifs
Protein Multimerization
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism

Substances

Cell Cycle Proteins
DNA-Binding Proteins
MAD2L2 protein, human
Mad2 Proteins
Phthalazines
Piperazines
Recombinant Proteins
SHLD1 protein, human
SHLD2 protein, human
SHLD3 protein, human
DNA
ATPases Associated with Diverse Cellular Activities
TRIP13 protein, human
Cisplatin
olaparib