RT Journal Article
SR Electronic
T1 Single-molecule live cell imaging of the Smc5/6 DNA repair complex
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.19.148106
DO 10.1101/2020.06.19.148106
A1 Etheridge, Thomas J.
A1 Villahermosa, Desiree
A1 Irmisch, Anja
A1 Watson, Adam T.
A1 Herbert, Alex
A1 Dang, Hung Q.
A1 Osborne, Mark A.
A1 Oliver, Antony W.
A1 Carr, Antony M.
A1 Murray, Johanne M.
YR 2020
UL http://biorxiv.org/content/early/2020/06/20/2020.06.19.148106.abstract
AB The Smc5/6 complex is involved in various DNA transactions and is best known for ensuring the fidelity of homologous recombination. We exploit 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 two 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. However, unlike a mutant attenuating chromatin association, a mutant that disrupts ssDNA binding results in highly elevated levels of gross chromosomal rearrangements during replication restart. This is consistent with a downstream function for ssDNA binding in regulating homologous recombination.Competing Interest StatementThe authors have declared no competing interest.