The PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeast

Protein-DNA complexes can impede DNA replication and cause replication fork collapse. Whilst it is known that homologous recombination is deployed in such instances to restart replication, it is unclear how a stalled fork transitions into a collapsed fork at which recombination proteins can load. Pr...

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Main Authors: Tamang, S, Kishkevich, A, Morrow, C, Osman, F, Jalan, M, Whitby, M
Format: Journal article
Published: eLife Sciences Publications 2019
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author Tamang, S
Kishkevich, A
Morrow, C
Osman, F
Jalan, M
Whitby, M
author_facet Tamang, S
Kishkevich, A
Morrow, C
Osman, F
Jalan, M
Whitby, M
author_sort Tamang, S
collection OXFORD
description Protein-DNA complexes can impede DNA replication and cause replication fork collapse. Whilst it is known that homologous recombination is deployed in such instances to restart replication, it is unclear how a stalled fork transitions into a collapsed fork at which recombination proteins can load. Previously we established assays in Schizosaccharomyces pombe for studying recombination induced by replication fork collapse at the site-specific protein-DNA barrier RTS1 (Nguyen et al., 2015). Here, we provide evidence that efficient recruitment/retention of two key recombination proteins (Rad51 and Rad52) to RTS1 depends on unloading of the polymerase sliding clamp PCNA from DNA by Elg1. We also show that, in the absence of Elg1, reduced recombination is partially suppressed by deleting fbh1 or, to a lesser extent, srs2, which encode known anti-recombinogenic DNA helicases. These findings suggest that PCNA unloading by Elg1 is necessary to limit Fbh1 and Srs2 activity, and thereby enable recombination to proceed.
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spelling oxford-uuid:0c7dc304-4288-400d-a1f0-21293d0f1b132022-03-26T09:35:20ZThe PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeastJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:0c7dc304-4288-400d-a1f0-21293d0f1b13Symplectic Elements at OxfordeLife Sciences Publications2019Tamang, SKishkevich, AMorrow, COsman, FJalan, MWhitby, MProtein-DNA complexes can impede DNA replication and cause replication fork collapse. Whilst it is known that homologous recombination is deployed in such instances to restart replication, it is unclear how a stalled fork transitions into a collapsed fork at which recombination proteins can load. Previously we established assays in Schizosaccharomyces pombe for studying recombination induced by replication fork collapse at the site-specific protein-DNA barrier RTS1 (Nguyen et al., 2015). Here, we provide evidence that efficient recruitment/retention of two key recombination proteins (Rad51 and Rad52) to RTS1 depends on unloading of the polymerase sliding clamp PCNA from DNA by Elg1. We also show that, in the absence of Elg1, reduced recombination is partially suppressed by deleting fbh1 or, to a lesser extent, srs2, which encode known anti-recombinogenic DNA helicases. These findings suggest that PCNA unloading by Elg1 is necessary to limit Fbh1 and Srs2 activity, and thereby enable recombination to proceed.
spellingShingle Tamang, S
Kishkevich, A
Morrow, C
Osman, F
Jalan, M
Whitby, M
The PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeast
title The PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeast
title_full The PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeast
title_fullStr The PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeast
title_full_unstemmed The PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeast
title_short The PCNA unloader Elg1 promotes recombination at collapsed replication forks in fission yeast
title_sort pcna unloader elg1 promotes recombination at collapsed replication forks in fission yeast
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