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Chromosomes; DNA damage and 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.
