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

Etheridge, Thomas J, Villahermosa, Desiree, Campillo-Funollet, Eduard, Herbert, Alex David, Irmisch, Anja, Watson, Adam T, Dang, Hung Q, Osborne, Mark A, Oliver, Antony W, Carr, Antony M, and others. (2021) Live-cell single-molecule tracking highlights requirements for stable Smc5/6 chromatin association in vivo. eLife, 10 . Article Number e68579. E-ISSN 2050-084X. (doi:10.7554/eLife.68579) (KAR id:90460)

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Official URL
https://doi.org/10.7554/eLife.68579

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: Article
DOI/Identification number: 10.7554/eLife.68579
Uncontrolled keywords: SMC proteins; single-molecule microscopy; DNA repair
Subjects: Q Science > QH Natural history > QH581.2 Cell Biology
Divisions: Divisions > Division of Computing, Engineering and Mathematical Sciences > School of Mathematics, Statistics and Actuarial Science
Depositing User: Amy Boaler
Date Deposited: 29 Sep 2021 13:15 UTC
Last Modified: 30 Sep 2021 09:48 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/90460 (The current URI for this page, for reference purposes)
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