A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome

Agnieszka Gambus, Frederick van Deursen, Dimitrios Polychronopoulos, Magdalena Foltman, Richard C. Jones, Ricky D. Edmondson, Arturo Calzada, Karim Labib (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    168 Citations (Scopus)

    Abstract

    The eukaryotic replisome is a crucial determinant of genome stability, but its structure is still poorly understood. We found previously that many regulatory proteins assemble around the MCM2-7 helicase at yeast replication forks to form the replisome progression complex (RPC), which might link MCM2-7 to other replisome components. Here, we show that the RPC associates with DNA polymerase alpha that primes each Okazaki fragment during lagging strand synthesis. Our data indicate that a complex of the GINS and Ctf4 components of the RPC is crucial to couple MCM2-7 to DNA polymerase alpha. Others have found recently that the Mrc1 subunit of RPCs binds DNA polymerase epsilon, which synthesises the leading strand at DNA replication forks. We show that cells lacking both Ctf4 and Mrc1 experience chronic activation of the DNA damage checkpoint during chromosome replication and do not complete the cell cycle. These findings indicate that coupling MCM2-7 to replicative polymerases is an important feature of the regulation of chromosome replication in eukaryotes, and highlight a key role for Ctf4 in this process.
    Original languageEnglish
    Pages (from-to)2992-3004
    Number of pages13
    JournalEMBO Journal
    Volume28
    Issue number19
    DOIs
    Publication statusPublished - 7 Oct 2009

    Fingerprint

    Minichromosome Maintenance Proteins
    DNA Polymerase I
    Chromosomes
    DNA Polymerase II
    Genomic Instability
    DNA
    Eukaryota
    DNA Replication
    Yeast
    DNA Damage
    Cell Cycle
    Genes
    Yeasts
    Chemical activation
    Cells
    Proteins

    Keywords

    • Chromosomal Proteins, Non-Histone
    • DNA Polymerase I
    • Cell Cycle Proteins
    • Nuclear Proteins
    • DNA-Binding Proteins
    • Saccharomyces cerevisiae Proteins
    • DNA
    • Protein Binding
    • Cell Cycle
    • Fungal Proteins
    • DNA Replication
    • Saccharomyces cerevisiae

    Cite this

    Gambus, A., van Deursen, F., Polychronopoulos, D., Foltman, M., Jones, R. C., Edmondson, R. D., ... Labib, K. (2009). A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome. EMBO Journal, 28(19), 2992-3004. https://doi.org/10.1038/emboj.2009.226
    Gambus, Agnieszka ; van Deursen, Frederick ; Polychronopoulos, Dimitrios ; Foltman, Magdalena ; Jones, Richard C. ; Edmondson, Ricky D. ; Calzada, Arturo ; Labib, Karim. / A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome. In: EMBO Journal. 2009 ; Vol. 28, No. 19. pp. 2992-3004.
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    title = "A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome",
    abstract = "The eukaryotic replisome is a crucial determinant of genome stability, but its structure is still poorly understood. We found previously that many regulatory proteins assemble around the MCM2-7 helicase at yeast replication forks to form the replisome progression complex (RPC), which might link MCM2-7 to other replisome components. Here, we show that the RPC associates with DNA polymerase alpha that primes each Okazaki fragment during lagging strand synthesis. Our data indicate that a complex of the GINS and Ctf4 components of the RPC is crucial to couple MCM2-7 to DNA polymerase alpha. Others have found recently that the Mrc1 subunit of RPCs binds DNA polymerase epsilon, which synthesises the leading strand at DNA replication forks. We show that cells lacking both Ctf4 and Mrc1 experience chronic activation of the DNA damage checkpoint during chromosome replication and do not complete the cell cycle. These findings indicate that coupling MCM2-7 to replicative polymerases is an important feature of the regulation of chromosome replication in eukaryotes, and highlight a key role for Ctf4 in this process.",
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    Gambus, A, van Deursen, F, Polychronopoulos, D, Foltman, M, Jones, RC, Edmondson, RD, Calzada, A & Labib, K 2009, 'A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome', EMBO Journal, vol. 28, no. 19, pp. 2992-3004. https://doi.org/10.1038/emboj.2009.226

    A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome. / Gambus, Agnieszka; van Deursen, Frederick; Polychronopoulos, Dimitrios; Foltman, Magdalena; Jones, Richard C.; Edmondson, Ricky D.; Calzada, Arturo; Labib, Karim (Lead / Corresponding author).

    In: EMBO Journal, Vol. 28, No. 19, 07.10.2009, p. 2992-3004.

    Research output: Contribution to journalArticle

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    AU - Gambus, Agnieszka

    AU - van Deursen, Frederick

    AU - Polychronopoulos, Dimitrios

    AU - Foltman, Magdalena

    AU - Jones, Richard C.

    AU - Edmondson, Ricky D.

    AU - Calzada, Arturo

    AU - Labib, Karim

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    N2 - The eukaryotic replisome is a crucial determinant of genome stability, but its structure is still poorly understood. We found previously that many regulatory proteins assemble around the MCM2-7 helicase at yeast replication forks to form the replisome progression complex (RPC), which might link MCM2-7 to other replisome components. Here, we show that the RPC associates with DNA polymerase alpha that primes each Okazaki fragment during lagging strand synthesis. Our data indicate that a complex of the GINS and Ctf4 components of the RPC is crucial to couple MCM2-7 to DNA polymerase alpha. Others have found recently that the Mrc1 subunit of RPCs binds DNA polymerase epsilon, which synthesises the leading strand at DNA replication forks. We show that cells lacking both Ctf4 and Mrc1 experience chronic activation of the DNA damage checkpoint during chromosome replication and do not complete the cell cycle. These findings indicate that coupling MCM2-7 to replicative polymerases is an important feature of the regulation of chromosome replication in eukaryotes, and highlight a key role for Ctf4 in this process.

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    KW - Protein Binding

    KW - Cell Cycle

    KW - Fungal Proteins

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    KW - Saccharomyces cerevisiae

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    Gambus A, van Deursen F, Polychronopoulos D, Foltman M, Jones RC, Edmondson RD et al. A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome. EMBO Journal. 2009 Oct 7;28(19):2992-3004. https://doi.org/10.1038/emboj.2009.226