elife-68579-v2.pdf (1.08 MB)
Live-cell single-molecule tracking highlights requirements for stable Smc5/6 chromatin association in vivo
Version 2 2023-06-12, 09:49
Version 1 2023-06-09, 23:38
journal contribution
posted on 2023-06-12, 09:49 authored by Thomas Etheridge, Desiree Villahermosa CaballeroDesiree Villahermosa Caballero, Eduard Campillo-Funollet, Alex David Herbert, Anja Irmisch, Adam WatsonAdam Watson, Hung Q Dang, Mark Osborne, Antony OliverAntony Oliver, Antony CarrAntony Carr, Jo MurrayThe 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 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.
Funding
How do Smc5/6 interactions with DNA coordinate replication and recombination?; G2119; MRC-MEDICAL RESEARCH COUNCIL; MR/P018955/1
Replication arrest, restart and genome instability; G1829; WELLCOME TRUST; 110047/Z/15/Z
History
Publication status
- Published
File Version
- Published version
Journal
eLifeISSN
2050-084XPublisher
eLife Sciences PublicationsExternal DOI
Volume
10Article number
a68579Department affiliated with
- Sussex Centre for Genome Damage Stability Publications
Full text available
- Yes
Peer reviewed?
- Yes
Legacy Posted Date
2021-04-19First Open Access (FOA) Date
2021-04-19First Compliant Deposit (FCD) Date
2021-04-16Usage metrics
Categories
No categories selectedKeywords
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC