Smc5/6 functions with Sgs1-Top3-Rmi1 to complete chromosome replication at natural pause sites

Sumedha Agashe, Chinnu Rose Joseph, Teresa Anne Clarisse Reyes, Demis Menolfi, Michele Giannattasio, Anja Waizenegger, Barnabas Szakal and Dana Branzei ()
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Sumedha Agashe: IFOM, the FIRC Institute of Molecular Oncology
Chinnu Rose Joseph: IFOM, the FIRC Institute of Molecular Oncology
Teresa Anne Clarisse Reyes: IFOM, the FIRC Institute of Molecular Oncology
Demis Menolfi: IFOM, the FIRC Institute of Molecular Oncology
Michele Giannattasio: IFOM, the FIRC Institute of Molecular Oncology
Anja Waizenegger: IFOM, the FIRC Institute of Molecular Oncology
Barnabas Szakal: IFOM, the FIRC Institute of Molecular Oncology
Dana Branzei: IFOM, the FIRC Institute of Molecular Oncology

Nature Communications, 2021, vol. 12, issue 1, 1-15

Abstract: Abstract Smc5/6 is essential for genome structural integrity by yet unknown mechanisms. Here we find that Smc5/6 co-localizes with the DNA crossed-strand processing complex Sgs1-Top3-Rmi1 (STR) at genomic regions known as natural pausing sites (NPSs) where it facilitates Top3 retention. Individual depletions of STR subunits and Smc5/6 cause similar accumulation of joint molecules (JMs) composed of reversed forks, double Holliday Junctions and hemicatenanes, indicative of Smc5/6 regulating Sgs1 and Top3 DNA processing activities. We isolate an intra-allelic suppressor of smc6-56 proficient in Top3 retention but affected in pathways that act complementarily with Sgs1 and Top3 to resolve JMs arising at replication termination. Upon replication stress, the smc6-56 suppressor requires STR and Mus81-Mms4 functions for recovery, but not Srs2 and Mph1 helicases that prevent maturation of recombination intermediates. Thus, Smc5/6 functions jointly with Top3 and STR to mediate replication completion and influences the function of other DNA crossed-strand processing enzymes at NPSs.

Date: 2021
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DOI: 10.1038/s41467-021-22217-w

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