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
Full text not available from this repository.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.