Replication-Coupled PCNA Unloading by the Elg1 Complex Occurs Genome-wide and Requires Okazaki Fragment Ligation

Takashi Kubota; Yuki Katou; Ryuichiro Nakato; Katsuhiko Shirahige; Anne Dunlop Donaldson

doi:10.1016/j.celrep.2015.06.066

Replication-Coupled PCNA Unloading by the Elg1 Complex Occurs Genome-wide and Requires Okazaki Fragment Ligation

Takashi Kubota, Yuki Katou, Ryuichiro Nakato, Katsuhiko Shirahige, Anne Dunlop Donaldson

Medical Sciences

The University of Tokyo

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76 Citations (Scopus)

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Abstract

The sliding clamp PCNA is a crucial component of the DNA replication machinery. Timely PCNA loading and unloading are central for genome integrity and must be strictly coordinated with other DNA processing steps during replication. Here, we show that the S. cerevisiae Elg1 replication factor C-like complex (Elg1-RLC) unloads PCNA genome-wide following Okazaki fragment ligation. In the absence of Elg1, PCNA is retained on chromosomes in the wake of replication forks, rather than at specific sites. Degradation of the Okazaki fragment ligase Cdc9 leads to PCNA accumulation on chromatin, similar to the accumulation caused by lack of Elg1. We demonstrate that Okazaki fragment ligation is the critical prerequisite for PCNA unloading, since Chlorella virus DNA ligase can substitute for Cdc9 in yeast and simultaneously promotes PCNA unloading. Our results suggest that Elg1-RLC acts as a general PCNA unloader and is dependent upon DNA ligation during chromosome replication.

Original language	English
Pages (from-to)	774-787
Number of pages	14
Journal	Cell Reports
Volume	12
Issue number	5
Early online date	23 Jul 2015
DOIs	https://doi.org/10.1016/j.celrep.2015.06.066
Publication status	Published - 4 Aug 2015

Bibliographical note

Open Access funded by Medical Research Council

Acknowledgments
We thank Dr Anja Bielinsky for plasmids and Dr. M.K. Raghuraman for a cdc9-1 strain. Alexander Lorenz (University of Aberdeen) provided valuable comments on the manuscript. This work was supported by Biotechnology and Biological Sciences Research Council (BBSRC) grant BB/K006304/1 to A.D., Medical Research Council Career Development Fellowship MR/L019698/1 to T.K., and MEXT Grant-in-Aid for Scientific Research on Innovative Areas to K.S.

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Replication-Coupled PCNA Unloading by the Elg1 Complex Occurs Genome-wide and Requires Okazaki Fragment Ligation
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/4.0/
Final published version, 4.07 MBLicence: CC BY

Anne Donaldson
Person: Academic
Takashi Kubota
Person: Academic, Academic Related - Research

Cite this

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title = "Replication-Coupled PCNA Unloading by the Elg1 Complex Occurs Genome-wide and Requires Okazaki Fragment Ligation",

abstract = "The sliding clamp PCNA is a crucial component of the DNA replication machinery. Timely PCNA loading and unloading are central for genome integrity and must be strictly coordinated with other DNA processing steps during replication. Here, we show that the S. cerevisiae Elg1 replication factor C-like complex (Elg1-RLC) unloads PCNA genome-wide following Okazaki fragment ligation. In the absence of Elg1, PCNA is retained on chromosomes in the wake of replication forks, rather than at specific sites. Degradation of the Okazaki fragment ligase Cdc9 leads to PCNA accumulation on chromatin, similar to the accumulation caused by lack of Elg1. We demonstrate that Okazaki fragment ligation is the critical prerequisite for PCNA unloading, since Chlorella virus DNA ligase can substitute for Cdc9 in yeast and simultaneously promotes PCNA unloading. Our results suggest that Elg1-RLC acts as a general PCNA unloader and is dependent upon DNA ligation during chromosome replication.",

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note = "Open Access funded by Medical Research Council Acknowledgments We thank Dr Anja Bielinsky for plasmids and Dr. M.K. Raghuraman for a cdc9-1 strain. Alexander Lorenz (University of Aberdeen) provided valuable comments on the manuscript. This work was supported by Biotechnology and Biological Sciences Research Council (BBSRC) grant BB/K006304/1 to A.D., Medical Research Council Career Development Fellowship MR/L019698/1 to T.K., and MEXT Grant-in-Aid for Scientific Research on Innovative Areas to K.S.",

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N1 - Open Access funded by Medical Research Council Acknowledgments We thank Dr Anja Bielinsky for plasmids and Dr. M.K. Raghuraman for a cdc9-1 strain. Alexander Lorenz (University of Aberdeen) provided valuable comments on the manuscript. This work was supported by Biotechnology and Biological Sciences Research Council (BBSRC) grant BB/K006304/1 to A.D., Medical Research Council Career Development Fellowship MR/L019698/1 to T.K., and MEXT Grant-in-Aid for Scientific Research on Innovative Areas to K.S.

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N2 - The sliding clamp PCNA is a crucial component of the DNA replication machinery. Timely PCNA loading and unloading are central for genome integrity and must be strictly coordinated with other DNA processing steps during replication. Here, we show that the S. cerevisiae Elg1 replication factor C-like complex (Elg1-RLC) unloads PCNA genome-wide following Okazaki fragment ligation. In the absence of Elg1, PCNA is retained on chromosomes in the wake of replication forks, rather than at specific sites. Degradation of the Okazaki fragment ligase Cdc9 leads to PCNA accumulation on chromatin, similar to the accumulation caused by lack of Elg1. We demonstrate that Okazaki fragment ligation is the critical prerequisite for PCNA unloading, since Chlorella virus DNA ligase can substitute for Cdc9 in yeast and simultaneously promotes PCNA unloading. Our results suggest that Elg1-RLC acts as a general PCNA unloader and is dependent upon DNA ligation during chromosome replication.

AB - The sliding clamp PCNA is a crucial component of the DNA replication machinery. Timely PCNA loading and unloading are central for genome integrity and must be strictly coordinated with other DNA processing steps during replication. Here, we show that the S. cerevisiae Elg1 replication factor C-like complex (Elg1-RLC) unloads PCNA genome-wide following Okazaki fragment ligation. In the absence of Elg1, PCNA is retained on chromosomes in the wake of replication forks, rather than at specific sites. Degradation of the Okazaki fragment ligase Cdc9 leads to PCNA accumulation on chromatin, similar to the accumulation caused by lack of Elg1. We demonstrate that Okazaki fragment ligation is the critical prerequisite for PCNA unloading, since Chlorella virus DNA ligase can substitute for Cdc9 in yeast and simultaneously promotes PCNA unloading. Our results suggest that Elg1-RLC acts as a general PCNA unloader and is dependent upon DNA ligation during chromosome replication.

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DO - 10.1016/j.celrep.2015.06.066

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JO - Cell Reports

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Replication-Coupled PCNA Unloading by the Elg1 Complex Occurs Genome-wide and Requires Okazaki Fragment Ligation

Abstract

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Anne Donaldson

Takashi Kubota

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