MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair
de Krijger, Inge; Föhr, Bastian; Pérez, Santiago Hernández; Vincendeau, Estelle; Serrat, Judit; Thouin, Alexander Marc; Susvirkar, Vivek; Lescale, Chloé; Paniagua, Inés; Hoekman, Liesbeth; Kaur, Simranjeet; Altelaar, Maarten; Deriano, Ludovic; Faesen, Alex C; Jacobs, Jacqueline J L
(2021) Nature Communications, volume 12, issue 1
(Article)
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
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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.
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Keywords: General Chemistry, General Biochemistry,Genetics and Molecular Biology, General Physics and Astronomy
ISSN: 2041-1723
Publisher: Nature Publishing Group
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 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).
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