Molecular mechanisms of assembly and TRIP13-mediated remodeling of the human Shieldin complex

Xie, Wei; Wang, Shengliu; Wang, Juncheng; de la Cruz, M. Jason; Xu, Guotai; Scaltriti, Maurizio; Patel, Dinshaw J.

doi:10.1073/pnas.2024512118

Title: Molecular mechanisms of assembly and TRIP13-mediated remodeling of the human Shieldin complex

Journal Article · Wed Feb 17 00:00:00 EST 2021 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.2024512118· OSTI ID:1768933

^[1]; Wang, Shengliu ^[1];

^[1];

^[1]; Xu, Guotai ^[1];

^[1];

^[1]

Memorial Sloan Kettering Cancer Center, New York, NY (United States)

The Shieldin complex, composed of REV7, SHLD1, SHLD2, and SHLD3, protects DNA double-strand breaks (DSBs) to promote nonhomologous end joining. The AAA⁺ ATPase TRIP13 remodels Shieldin to regulate DNA repair pathway choice. Here we report crystal structures of human SHLD3–REV7 binary and fused SHLD2–SHLD3–REV7 ternary complexes, revealing that assembly of Shieldin requires fused SHLD2–SHLD3 induced conformational heterodimerization of open (O-REV7) and closed (C-REV7) forms of REV7. We also report the cryogenic electron microscopy (cryo-EM) structures of the ATPγS-bound fused SHLD2–SHLD3–REV7–TRIP13 complexes, uncovering the principles underlying the TRIP13-mediated disassembly mechanism of the Shieldin complex. We demonstrate that the N terminus of REV7 inserts into the central channel of TRIP13, setting the stage for pulling the unfolded N-terminal peptide of C-REV7 through the central TRIP13 hexameric channel. The primary interface involves contacts between the safety-belt segment of C-REV7 and a conserved and negatively charged loop of TRIP13. This process is mediated by ATP hydrolysis-triggered rotatory motions of the TRIP13 ATPase, thereby resulting in the disassembly of the Shieldin complex.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC); Memorial Sloan Kettering Cancer Center; National Institutes of Health (NIH)

Grant/Contract Number:: AC02‐06CH11357; P30-CA016086; P30 GM124165; S10 RR029205

OSTI ID:: 1768933

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 118, Issue 8; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
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SHLD2-SHLD3-REV7-TRIP13 complex

Title: Molecular mechanisms of assembly and TRIP13-mediated remodeling of the human Shieldin complex

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References (55)

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