1. MAD2L2 dimerization and TRIP13 control shieldin activity in DNA repair.
- Author
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de Krijger I, Föhr B, Pérez SH, Vincendeau E, Serrat J, Thouin AM, Susvirkar V, Lescale C, Paniagua I, Hoekman L, Kaur S, Altelaar M, Deriano L, Faesen AC, and Jacobs JJL
- Subjects
- ATPases Associated with Diverse Cellular Activities chemistry, ATPases Associated with Diverse Cellular Activities metabolism, Animals, Binding Sites, Cell Cycle Proteins chemistry, Cell Cycle Proteins metabolism, Cell Line, Cell Line, Tumor, Cisplatin pharmacology, DNA chemistry, DNA metabolism, DNA Breaks, Double-Stranded, DNA-Binding Proteins chemistry, DNA-Binding Proteins metabolism, Fibroblasts cytology, Fibroblasts drug effects, Fibroblasts metabolism, Gene Expression, HEK293 Cells, HeLa Cells, Humans, Mad2 Proteins chemistry, 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, ATPases Associated with Diverse Cellular Activities genetics, Cell Cycle Proteins genetics, DNA genetics, DNA Repair, DNA-Binding Proteins genetics, Mad2 Proteins genetics
- 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., (© 2021. The Author(s).)
- Published
- 2021
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