Mcm10 associates with the loaded DNA helicase at replication origins and defines a novel step in its activation

Frederick van Deursen, Sugopa Sengupta, Giacomo De Piccoli, Alberto Sanchez-Diaz, Karim Labib (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    87 Citations (Scopus)

    Abstract

    Mcm10 is essential for chromosome replication in eukaryotic cells and was previously thought to link the Mcm2-7 DNA helicase at replication forks to DNA polymerase alpha. Here, we show that yeast Mcm10 interacts preferentially with the fraction of the Mcm2-7 helicase that is loaded in an inactive form at origins of DNA replication, suggesting a role for Mcm10 during the initiation of chromosome replication, but Mcm10 is not a stable component of the replisome subsequently. Studies with budding yeast and human cells indicated that Mcm10 chaperones the catalytic subunit of polymerase alpha and preserves its stability. We used a novel degron allele to inactivate Mcm10 efficiently and this blocked the initiation of chromosome replication without causing degradation of DNA polymerase alpha. Strikingly, the other essential helicase subunits Cdc45 and GINS were still recruited to Mcm2-7 when cells entered S-phase without Mcm10, but origin unwinding was blocked. These findings indicate that Mcm10 is required for a novel step during activation of the Cdc45-MCM-GINS helicase at DNA replication origins.
    Original languageEnglish
    Pages (from-to)2195-2206
    Number of pages12
    JournalEMBO Journal
    Volume31
    Issue number9
    DOIs
    Publication statusPublished - 2 May 2012

    Keywords

    • Yeasts
    • Chromosomal Proteins, Non-Histone
    • DNA, Fungal
    • Cell Cycle Proteins
    • DNA Helicases
    • Fungal Proteins
    • DNA Replication
    • Antigens, CD45

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