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      The small subunit of DNA polymerase D (DP1) associates with GINS–GAN complex of the thermophilic archaea in Thermococcus sp. 4557

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          Abstract

          The eukaryotic GINS, Cdc45, and minichromosome maintenance proteins form an essential complex that moves with the DNA replication fork. The GINS protein complex has also been reported to associate with DNA polymerase. In archaea, the third domain of life, DNA polymerase D (PolD) is essential for DNA replication, and the genes encoding PolDs exist only in the genomes of archaea. The archaeal GAN ( GINS‐ associated nuclease) is believed to be a homolog of the eukaryotic Cdc45. In this study, we found that the Thermococcus sp. 4557 DP1 (small subunit of PolD) interacted with GINS15 in vitro, and the 3′–5′ exonuclease activity of DP1 was inhibited by GINS15. We also demonstrated that the GAN, GINS15, and DP1 proteins interact to form a complex adapting a GAN–GINS15–DP1 order. The results of this study imply that the complex constitutes a core of the DNA replisome in archaea.

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          Most cited references47

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          GINS maintains association of Cdc45 with MCM in replisome progression complexes at eukaryotic DNA replication forks.

          The components of the replisome that preserve genomic stability by controlling the progression of eukaryotic DNA replication forks are poorly understood. Here, we show that the GINS (go ichi ni san) complex allows the MCM (minichromosome maintenance) helicase to interact with key regulatory proteins in large replisome progression complexes (RPCs) that are assembled during initiation and disassembled at the end of S phase. RPC components include the essential initiation and elongation factor, Cdc45, the checkpoint mediator Mrc1, the Tof1-Csm3 complex that allows replication forks to pause at protein-DNA barriers, the histone chaperone FACT (facilitates chromatin transcription) and Ctf4, which helps to establish sister chromatid cohesion. RPCs also interact with Mcm10 and topoisomerase I. During initiation, GINS is essential for a specific subset of RPC proteins to interact with MCM. GINS is also important for the normal progression of DNA replication forks, and we show that it is required after initiation to maintain the association between MCM and Cdc45 within RPCs.
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            Eukaryotic DNA Replication Fork.

            This review focuses on the biogenesis and composition of the eukaryotic DNA replication fork, with an emphasis on the enzymes that synthesize DNA and repair discontinuities on the lagging strand of the replication fork. Physical and genetic methodologies aimed at understanding these processes are discussed. The preponderance of evidence supports a model in which DNA polymerase ε (Pol ε) carries out the bulk of leading strand DNA synthesis at an undisturbed replication fork. DNA polymerases α and δ carry out the initiation of Okazaki fragment synthesis and its elongation and maturation, respectively. This review also discusses alternative proposals, including cellular processes during which alternative forks may be utilized, and new biochemical studies with purified proteins that are aimed at reconstituting leading and lagging strand DNA synthesis separately and as an integrated replication fork.
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              Isolation of the Cdc45/Mcm2-7/GINS (CMG) complex, a candidate for the eukaryotic DNA replication fork helicase.

              The protein Cdc45 plays a critical but poorly understood role in the initiation and elongation stages of eukaryotic DNA replication. To study Cdc45's function in DNA replication, we purified Cdc45 protein from Drosophila embryo extracts by a combination of traditional and immunoaffinity chromatography steps and found that the protein exists in a stable, high-molecular-weight complex with the Mcm2-7 hexamer and the GINS tetramer. The purified Cdc45/Mcm2-7/GINS complex is associated with an active ATP-dependent DNA helicase function. RNA interference knock-down experiments targeting the GINS and Cdc45 components establish that the proteins are required for the S phase transition in Drosophila cells. The data suggest that this complex forms the core helicase machinery for eukaryotic DNA replication.
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                Author and article information

                Contributors
                lizhuo@tio.org.cn
                Journal
                Microbiologyopen
                Microbiologyopen
                10.1002/(ISSN)2045-8827
                MBO3
                MicrobiologyOpen
                John Wiley and Sons Inc. (Hoboken )
                2045-8827
                08 May 2019
                September 2019
                : 8
                : 9 ( doiID: 10.1002/mbo3.v8.9 )
                : e00848
                Affiliations
                [ 1 ] College of Ocean and Earth Sciences Xiamen University Xiamen Fujian China
                [ 2 ] Third Institute of Oceanography, Ministry of Natural Resources of China Xiamen Fujian China
                [ 3 ] School of Medicine Guangxi University Nanning Guangxi China
                [ 4 ] State Key Laboratory of Microbial Technology Shandong University Qingdao Shandong China
                Author notes
                [*] [* ] Correspondence

                Zhuo Li, Third Institute of Oceanography, Ministry of Natural Resources of China, Xiamen, Fujian, China.

                Email: lizhuo@ 123456tio.org.cn

                Author information
                https://orcid.org/0000-0003-3453-5903
                Article
                MBO3848
                10.1002/mbo3.848
                6741145
                31069963
                af2b7b59-746d-4c73-a50a-4d87efa0ddc7
                © 2019 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 12 March 2019
                : 26 March 2019
                : 27 March 2019
                Page count
                Figures: 9, Tables: 1, Pages: 15, Words: 7470
                Funding
                Funded by: National Natural Science Foundation of China
                Award ID: 31500027
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                mbo3848
                September 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.9 mode:remove_FC converted:12.09.2019

                Microbiology & Virology
                archaea,dna replication,gan,gins,pold
                Microbiology & Virology
                archaea, dna replication, gan, gins, pold

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