TY - JOUR
T1 - Structural and functional characterization of the Spo11 core complex
AU - Claeys Bouuaert, Corentin
AU - Tischfield, Sam E.
AU - Pu, Stephen
AU - Mimitou, Eleni P.
AU - Arias-Palomo, Ernesto
AU - Berger, James M.
AU - Keeney, Scott
N1 - Funding Information:
We thank M. Brendel and the Molecular Cytology core facility at MSKCC for performing the AFM experiments. We thank R. Hendrickson, E. Chang and the Microchemistry and Proteomics core facility at MSKCC for assistance with the XL–MS experiments. We thank K. Liu (S.K. laboratory) for discussions. MSKCC core facilities are supported by National Cancer Institute (NCI) Cancer Center support grant no. P30 CA08748. We thank E. Folta-Stogniew and the Biophysics Resource of Keck Facility at Yale University for the SEC–MALS experiments. The SEC–LS/UV/RI instrumentation was supported by NIH Award Number 1S10RR023748-01. E.P.M. was supported in part by a Helen Hay Whitney Foundation fellowship. Work in the S.K. laboratory was supported principally by the Howard Hughes Medical Institute and in part by NIH grant no. R35 GM118092 (S.K.). Work in the J.M.B. laboratory was funded by NCI grant no. R01-CA0777373 (J.M.B.). C.C.B. was supported in part by funding from the European Research Council under the European Union’s Horizon 2020 research and innovation program (European Research Council grant agreement no. 802525) and from the Fonds National de la Recherche Scientifique (FNRS MIS-Ulysse grant no. F.6002.20) (C.C.B.).
Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2021/1
Y1 - 2021/1
N2 - Spo11, which makes DNA double-strand breaks (DSBs) that are essential for meiotic recombination, has long been recalcitrant to biochemical study. We provide molecular analysis of Saccharomycescerevisiae Spo11 purified with partners Rec102, Rec104 and Ski8. Rec102 and Rec104 jointly resemble the B subunit of archaeal topoisomerase VI, with Rec104 occupying a position similar to the Top6B GHKL-type ATPase domain. Unexpectedly, the Spo11 complex is monomeric (1:1:1:1 stoichiometry), consistent with dimerization controlling DSB formation. Reconstitution of DNA binding reveals topoisomerase-like preferences for duplex–duplex junctions and bent DNA. Spo11 also binds noncovalently but with high affinity to DNA ends mimicking cleavage products, suggesting a mechanism to cap DSB ends. Mutations that reduce DNA binding in vitro attenuate DSB formation, alter DSB processing and reshape the DSB landscape in vivo. Our data reveal structural and functional similarities between the Spo11 core complex and Topo VI, but also highlight differences reflecting their distinct biological roles.
AB - Spo11, which makes DNA double-strand breaks (DSBs) that are essential for meiotic recombination, has long been recalcitrant to biochemical study. We provide molecular analysis of Saccharomycescerevisiae Spo11 purified with partners Rec102, Rec104 and Ski8. Rec102 and Rec104 jointly resemble the B subunit of archaeal topoisomerase VI, with Rec104 occupying a position similar to the Top6B GHKL-type ATPase domain. Unexpectedly, the Spo11 complex is monomeric (1:1:1:1 stoichiometry), consistent with dimerization controlling DSB formation. Reconstitution of DNA binding reveals topoisomerase-like preferences for duplex–duplex junctions and bent DNA. Spo11 also binds noncovalently but with high affinity to DNA ends mimicking cleavage products, suggesting a mechanism to cap DSB ends. Mutations that reduce DNA binding in vitro attenuate DSB formation, alter DSB processing and reshape the DSB landscape in vivo. Our data reveal structural and functional similarities between the Spo11 core complex and Topo VI, but also highlight differences reflecting their distinct biological roles.
UR - http://www.scopus.com/inward/record.url?scp=85098790244&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85098790244&partnerID=8YFLogxK
U2 - 10.1038/s41594-020-00534-w
DO - 10.1038/s41594-020-00534-w
M3 - Article
C2 - 33398171
AN - SCOPUS:85098790244
SN - 1545-9993
VL - 28
SP - 92
EP - 102
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
IS - 1
ER -