MAD2L2 Dimerization and TRIP13 Control Shieldin Activity in DNA Repair

Overview

Journal Nat Commun

Specialty Biology

Date 2021 Sep 15

PMID 34521823

Citations 21

Authors

Inge de Krijger

Bastian Fohr

Santiago Hernandez Perez

Estelle Vincendeau

Judit Serrat

Alexander Marc Thouin

Vivek Susvirkar

Chloe Lescale

Ines Paniagua

Liesbeth Hoekman

Simranjeet Kaur

Maarten Altelaar

Ludovic Deriano

Alex C Faesen

Jacqueline J L Jacobs

Affiliations

Soon will be listed here.

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.

Citing Articles

POGZ targeted by LINC01355/miR-27b-3p retards thyroid cancer progression via interplaying with MAD2L2.

Lu J, Zhou X, Zhu H, Zou M, Liu L, Li X 3 Biotech. 2025; 15(4):79.

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TRIP13 protects pancreatic cancer cells against intrinsic and therapy-induced DNA replication stress.

Anand J, Droby G, Joseph S, Patel U, Zhang X, Klomp J bioRxiv. 2025; .

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REV7: a small but mighty regulator of genome maintenance and cancer development.

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The RING Finger E3 Ligase RNF25 Protects DNA Replication Forks Independently of its Canonical Roles in Ubiquitin Signaling.

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References

Howard S, Ceppi I, Anand R, Geiger R, Cejka P . The internal region of CtIP negatively regulates DNA end resection. Nucleic Acids Res. 2020; 48(10):5485-5498. PMC: 7261161. DOI: 10.1093/nar/gkaa273. View

Gunn A, Stark J . I-SceI-based assays to examine distinct repair outcomes of mammalian chromosomal double strand breaks. Methods Mol Biol. 2012; 920:379-91. DOI: 10.1007/978-1-61779-998-3_27. View

Rizzo A, Vassel F, Chatterjee N, DSouza S, Li Y, Hao B . Rev7 dimerization is important for assembly and function of the Rev1/Polζ translesion synthesis complex. Proc Natl Acad Sci U S A. 2018; 115(35):E8191-E8200. PMC: 6126741. DOI: 10.1073/pnas.1801149115. View

Vader G . Pch2(TRIP13): controlling cell division through regulation of HORMA domains. Chromosoma. 2015; 124(3):333-9. DOI: 10.1007/s00412-015-0516-y. View

Boboila C, Yan C, Wesemann D, Jankovic M, Wang J, Manis J . Alternative end-joining catalyzes class switch recombination in the absence of both Ku70 and DNA ligase 4. J Exp Med. 2010; 207(2):417-27. PMC: 2822597. DOI: 10.1084/jem.20092449. View

Yan C, Boboila C, Souza E, Franco S, Hickernell T, Murphy M . IgH class switching and translocations use a robust non-classical end-joining pathway. Nature. 2007; 449(7161):478-82. DOI: 10.1038/nature06020. View

Boersma V, Moatti N, Segura-Bayona S, Peuscher M, van der Torre J, Wevers B . MAD2L2 controls DNA repair at telomeres and DNA breaks by inhibiting 5' end resection. Nature. 2015; 521(7553):537-540. PMC: 4481296. DOI: 10.1038/nature14216. View

Sarangi P, Clairmont C, Galli L, Moreau L, DAndrea A . p31 promotes homologous recombination by inactivating REV7 through the TRIP13 ATPase. Proc Natl Acad Sci U S A. 2020; 117(43):26795-26803. PMC: 7604461. DOI: 10.1073/pnas.2008830117. View

Setiaputra D, Durocher D . Shieldin - the protector of DNA ends. EMBO Rep. 2019; 20(5). PMC: 6501030. DOI: 10.15252/embr.201847560. View

10.

Gao S, Feng S, Ning S, Liu J, Zhao H, Xu Y . An OB-fold complex controls the repair pathways for DNA double-strand breaks. Nat Commun. 2018; 9(1):3925. PMC: 6156606. DOI: 10.1038/s41467-018-06407-7. View

11.

Malik R, Kopylov M, Gomez-Llorente Y, Jain R, Johnson R, Prakash L . Structure and mechanism of B-family DNA polymerase ζ specialized for translesion DNA synthesis. Nat Struct Mol Biol. 2020; 27(10):913-924. PMC: 7554088. DOI: 10.1038/s41594-020-0476-7. View

12.

Xu G, Chapman J, Brandsma I, Yuan J, Mistrik M, Bouwman P . REV7 counteracts DNA double-strand break resection and affects PARP inhibition. Nature. 2015; 521(7553):541-544. PMC: 4671316. DOI: 10.1038/nature14328. View

13.

Harmsen T, Klaasen S, van de Vrugt H, Te Riele H . DNA mismatch repair and oligonucleotide end-protection promote base-pair substitution distal from a CRISPR/Cas9-induced DNA break. Nucleic Acids Res. 2018; 46(6):2945-2955. PMC: 5888797. DOI: 10.1093/nar/gky076. View

14.

Lescale C, Abramowski V, Bedora-Faure M, Murigneux V, Vera G, Roth D . RAG2 and XLF/Cernunnos interplay reveals a novel role for the RAG complex in DNA repair. Nat Commun. 2016; 7:10529. PMC: 4740868. DOI: 10.1038/ncomms10529. View

15.

Tang J, Cho N, Cui G, Manion E, Shanbhag N, Botuyan M . Acetylation limits 53BP1 association with damaged chromatin to promote homologous recombination. Nat Struct Mol Biol. 2013; 20(3):317-25. PMC: 3594358. DOI: 10.1038/nsmb.2499. View

16.

Findlay S, Heath J, Luo V, Malina A, Morin T, Coulombe Y . SHLD2/FAM35A co-operates with REV7 to coordinate DNA double-strand break repair pathway choice. EMBO J. 2018; 37(18). PMC: 6138439. DOI: 10.15252/embj.2018100158. View

17.

Hara K, Shimizu T, Unzai S, Akashi S, Sato M, Hashimoto H . Purification, crystallization and initial X-ray diffraction study of human REV7 in complex with a REV3 fragment. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010; 65(Pt 12):1302-5. PMC: 2802887. DOI: 10.1107/S1744309109046181. View

18.

Ye Q, Kim D, Dereli I, Rosenberg S, Hagemann G, Herzog F . The AAA+ ATPase TRIP13 remodels HORMA domains through N-terminal engagement and unfolding. EMBO J. 2017; 36(16):2419-2434. PMC: 5556265. DOI: 10.15252/embj.201797291. View

19.

Dai Y, Zhang F, Wang L, Shan S, Gong Z, Zhou Z . Structural basis for shieldin complex subunit 3-mediated recruitment of the checkpoint protein REV7 during DNA double-strand break repair. J Biol Chem. 2019; 295(1):250-262. PMC: 6952594. DOI: 10.1074/jbc.RA119.011464. View

20.

Zimmermann M, Murina O, Reijns M, Agathanggelou A, Challis R, Tarnauskaite Z . CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions. Nature. 2018; 559(7713):285-289. PMC: 6071917. DOI: 10.1038/s41586-018-0291-z. View