Live-cell single-molecule tracking highlights requirements for stable Smc5/6 chromatin association in vivo

Etheridge, Thomas J and Villahermosa, Desiree and Campillo-Funollet, Eduard and Herbert, Alex David and Irmisch, Anja and Watson, Adam T and Dang, Hung Q and Osborne, Mark A and Oliver, Antony W and Carr, Antony M and Murray, Johanne M (2021) Live-cell single-molecule tracking highlights requirements for stable Smc5/6 chromatin association in vivo. eLife, 10: e68579. ISSN 2050-084X

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Abstract

The essential Smc5/6 complex is required in response to replication stress and is best known for ensuring the fidelity of homologous recombination. Using single-molecule tracking in live fission yeast to investigate Smc5/6 chromatin association, we show that Smc5/6 is chromatin associated in unchallenged cells and this depends on the non-SMC protein Nse6. We define a minimum of two Nse6-dependent sub-pathways, one of which requires the BRCT-domain protein Brc1. Using defined mutants in genes encoding the core Smc5/6 complex subunits, we show that the Nse3 double-stranded DNA binding activity and the arginine fingers of the two Smc5/6 ATPase binding sites are critical for chromatin association. Interestingly, disrupting the single-stranded DNA (ssDNA) binding activity at the hinge region does not prevent chromatin association but leads to elevated levels of gross chromosomal rearrangements during replication restart. This is consistent with a downstream function for ssDNA binding in regulating homologous recombination.

Item Type:
Journal Article
Journal or Publication Title:
eLife
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/1300/1300
Subjects:
?? general biochemistry,genetics and molecular biologygeneral medicinegeneral immunology and microbiologygeneral neurosciencebiochemistry, genetics and molecular biology(all)medicine(all)immunology and microbiology(all)neuroscience(all) ??
ID Code:
184758
Deposited By:
Deposited On:
13 Feb 2023 10:55
Refereed?:
Yes
Published?:
Published
Last Modified:
16 Jul 2024 11:58