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

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

Division of Psychosocial Research and Epidemiology, The Netherlands Cancer Institute - Antoni van Leeuwenhoek Hospital, Amsterdam, the Netherlands.
Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute)
Genome Integrity

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Article number	5421
Number of pages	15
Journal	Nature Communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-25724-y
Publication status	Published - Dec 2021

Access to Document

10.1038/s41467-021-25724-yLicence: CC BY

s41467-021-25724-yFinal published version, 2.3 MBLicence: CC BY

Cite this

de Krijger, I., Föhr, B., Pérez, S. H., Vincendeau, E., Serrat, J., Thouin, A. M., Susvirkar, V., Lescale, C., Paniagua, I., Hoekman, L., Kaur, S., Altelaar, M., Deriano, L., Faesen, A. C., & Jacobs, J. J. L. (2021). MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair. Nature Communications, 12(1), Article 5421. https://doi.org/10.1038/s41467-021-25724-y

@article{8fc813a5f54443589ce650c856aa8477,

title = "MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair",

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.",

author = "{de Krijger}, Inge and Bastian F{\"o}hr and P{\'e}rez, {Santiago Hern{\'a}ndez} and Estelle Vincendeau and Judit Serrat and Thouin, {Alexander Marc} and Vivek Susvirkar and Chlo{\'e} Lescale and In{\'e}s Paniagua and Liesbeth Hoekman and Simranjeet Kaur and Maarten Altelaar and Ludovic Deriano and Faesen, {Alex C} and Jacobs, {Jacqueline J L}",

note = "Funding Information: We thank T. de Lange for the Trf2ts allele and Trf2flox/−;p53−/− MEFs, D. Durocher for BRCA1−/− RPE, R. Greenberg for U2OS-DSB reporter cells with inducible mCherry-LacI-Fok1 nuclease, G. Vader for HeLa cells expressing doxycycline-inducible GFP-tagged TRIP13EQ and TRIP13 expression constructs, B. de Wolf and G. Kops for TRIP13 KO HeLa, G. Legube for AID-DIvA U2OS cells, J. Stark for DR-GFP U2OS cells, C. Choudhary for GFP-RINN1 (SHLD3) and RINN2 (SHLD2) expression constructs, M. Barazas for pX330p-sgNT, B. van den Broek for customized macros for image analysis and R. Medema and B. Rowland labs for discussions. This work was supported by project grant 10999/2017-1 from the Dutch Cancer Society (KWF) to J.J.L.J., by the Institut Pasteur to L.D., by the Institut National Du Cancer (INCA_13852) to L.D., by the Ligue Nationale Contre le Cancer (Equipe labellis{\'e}e 2019) to L.D. The Proteomics Facility of the Netherlands Cancer Institute was supported by the X-omics Initiative, partially funded by the Dutch Research Council (NWO)(project 184.034.019). E.V. received funding from Universit{\'e} de Paris, Sorbonne Paris Cit{\'e}. A.C.F. is grateful for generous core funding by the Max Planck Society. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

year = "2021",

month = dec,

doi = "10.1038/s41467-021-25724-y",

language = "English",

volume = "12",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Research",

number = "1",

}

TY - JOUR

T1 - MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair

AU - de Krijger, Inge

AU - Föhr, Bastian

AU - Pérez, Santiago Hernández

AU - Vincendeau, Estelle

AU - Serrat, Judit

AU - Thouin, Alexander Marc

AU - Susvirkar, Vivek

AU - Lescale, Chloé

AU - Paniagua, Inés

AU - Hoekman, Liesbeth

AU - Kaur, Simranjeet

AU - Altelaar, Maarten

AU - Deriano, Ludovic

AU - Faesen, Alex C

AU - Jacobs, Jacqueline J L

N1 - Funding Information: We thank T. de Lange for the Trf2ts allele and Trf2flox/−;p53−/− MEFs, D. Durocher for BRCA1−/− RPE, R. Greenberg for U2OS-DSB reporter cells with inducible mCherry-LacI-Fok1 nuclease, G. Vader for HeLa cells expressing doxycycline-inducible GFP-tagged TRIP13EQ and TRIP13 expression constructs, B. de Wolf and G. Kops for TRIP13 KO HeLa, G. Legube for AID-DIvA U2OS cells, J. Stark for DR-GFP U2OS cells, C. Choudhary for GFP-RINN1 (SHLD3) and RINN2 (SHLD2) expression constructs, M. Barazas for pX330p-sgNT, B. van den Broek for customized macros for image analysis and R. Medema and B. Rowland labs for discussions. This work was supported by project grant 10999/2017-1 from the Dutch Cancer Society (KWF) to J.J.L.J., by the Institut Pasteur to L.D., by the Institut National Du Cancer (INCA_13852) to L.D., by the Ligue Nationale Contre le Cancer (Equipe labellisée 2019) to L.D. The Proteomics Facility of the Netherlands Cancer Institute was supported by the X-omics Initiative, partially funded by the Dutch Research Council (NWO)(project 184.034.019). E.V. received funding from Université de Paris, Sorbonne Paris Cité. A.C.F. is grateful for generous core funding by the Max Planck Society. Publisher Copyright: © 2021, The Author(s).

PY - 2021/12

Y1 - 2021/12

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85115222105&partnerID=8YFLogxK

U2 - 10.1038/s41467-021-25724-y

DO - 10.1038/s41467-021-25724-y

M3 - Article

C2 - 34521823

SN - 2041-1723

VL - 12

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 5421

ER -

MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair

Abstract

Access to Document

Fingerprint

Cite this