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Ctf18-RFC and DNA Pol ϵ form a stable leading strand polymerase/clamp loader complex required for normal and perturbed DNA replication

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Stokes, Katherine, Winczura, Alicja, Song, Boyuan, De Piccoli, Giacomo and Grabarczyk, Daniel B. (2020) Ctf18-RFC and DNA Pol ϵ form a stable leading strand polymerase/clamp loader complex required for normal and perturbed DNA replication. Nucleic Acids Research, 48 (14). pp. 8128-8145. doi:10.1093/nar/gkaa541

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Official URL: https://doi.org/10.1093/nar/gkaa541

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Abstract

The eukaryotic replisome must faithfully replicate DNA and cope with replication fork blocks and stalling, while simultaneously promoting sister chromatid cohesion. Ctf18-RFC is an alternative PCNA loader that links all these processes together by an unknown mechanism. Here, we use integrative structural biology combined with yeast genetics and biochemistry to highlight the specific functions that Ctf18-RFC plays within the leading strand machinery via an interaction with the catalytic domain of DNA Pol ϵ. We show that a large and unusually flexible interface enables this interaction to occur constitutively throughout the cell cycle and regardless of whether forks are replicating or stalled. We reveal that, by being anchored to the leading strand polymerase, Ctf18-RFC can rapidly signal fork stalling to activate the S phase checkpoint. Moreover, we demonstrate that, independently of checkpoint signaling or chromosome cohesion, Ctf18-RFC functions in parallel to Chl1 and Mrc1 to protect replication forks and cell viability. [Abstract copyright: © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.]

Item Type: Journal Article
Subjects: Q Science > QH Natural history > QH426 Genetics
Divisions: Faculty of Medicine > Warwick Medical School > Biomedical Sciences
Faculty of Medicine > Warwick Medical School
SWORD Depositor: Library Publications Router
Library of Congress Subject Headings (LCSH): DNA replication, DNA polymerases, Proliferating cell nuclear antigen
Journal or Publication Title: Nucleic Acids Research
Publisher: Oxford University Press
ISSN: 0305-1048
Official Date: 25 June 2020
Dates:
DateEvent
25 June 2020Published
16 June 2020Accepted
Date of first compliant deposit: 2 September 2020
Volume: 48
Number: 14
Page Range: pp. 8128-8145
DOI: 10.1093/nar/gkaa541
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
C44595/A16326Cancer Research UKhttp://dx.doi.org/10.13039/501100000289
GR 5152/3-1[DFG] Deutsche Forschungsgemeinschafthttp://dx.doi.org/10.13039/501100001659

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