Live-cell single-molecule tracking highlights requirements for stable Smc5/6 chromatin association in vivo
| Autor: | Anja Irmisch, Eduard Campillo-Funollet, Alex Herbert, Mark A. Osborne, Hung Quang Dang, Johanne M. Murray, Antony W. Oliver, Antony M. Carr, Adam T. Watson, Thomas J. Etheridge, Desirée Villahermosa |
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| Rok vydání: | 2021 |
| Předmět: |
DNA repair
QH301-705.5 Science Mutant Cell Cycle Proteins ATPase binding General Biochemistry Genetics and Molecular Biology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Schizosaccharomyces single-molecule microscopy Biology (General) Gene QH581.2 030304 developmental biology 0303 health sciences General Immunology and Microbiology Chemistry General Neuroscience SMC protein Genetics and Genomics General Medicine Chromosomes and Gene Expression Chromatin Single Molecule Imaging Cell biology SMC proteins Medicine Schizosaccharomyces pombe Proteins Homologous recombination 030217 neurology & neurosurgery DNA Research Article S. pombe |
| Zdroj: | eLife eLife, Vol 10 (2021) |
| ISSN: | 2050-084X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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