DNA synthesis at individual replication forks requires the essential initiation factor Cdc45p

JA Tercero, K Labib, JFX Diffley

    Research output: Contribution to journalArticle

    146 Citations (Scopus)

    Abstract

    Cdc45p assembles at replication origins before initiation and is required for origin firing in Saccharomyces cerevisiae. A heat-inducible cdc45 degron mutant was constructed that promotes rapid degradation of Cdc45p at the restrictive temperature, Consistent with a role in initiation, loss of Cdc45p in G(I) prevents all detectable DNA replication without preventing subsequent entry into mitosis, Loss of Cdc45p activity during S-phase blocks S-phase completion but not activation of replication checkpoints. Using density substitution, me show that after allowing replication fork establishment, Cdc45p inactivation prevents the subsequent progression of individual replication forks, This provides the first direct functional evidence that Cdc45p plays an essential role during elongation. Thus, like the large T antigen in SV40 replication, Cdc45p plays a central role in both initiation and elongation phases of chromosomal DNA replication.

    Original languageEnglish
    Pages (from-to)2082-2093
    Number of pages12
    JournalEMBO Journal
    Volume19
    Issue number9
    DOIs
    Publication statusPublished - 2 May 2000

    Keywords

    • S-phase
    • ORIGIN RECOGNITION COMPLEX
    • POLYMERASE-ALPHA
    • pre-replicative complex
    • S-PHASE
    • cell cycle
    • CEREVISIAE CHROMOSOME-VI
    • MCM PROTEINS
    • BUDDING YEAST
    • CELL-CYCLE
    • HELICASE ACTIVITY
    • DNA polymerase
    • MINICHROMOSOME MAINTENANCE
    • DNA replication
    • SACCHAROMYCES-CEREVISIAE

    Cite this

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    title = "DNA synthesis at individual replication forks requires the essential initiation factor Cdc45p",
    abstract = "Cdc45p assembles at replication origins before initiation and is required for origin firing in Saccharomyces cerevisiae. A heat-inducible cdc45 degron mutant was constructed that promotes rapid degradation of Cdc45p at the restrictive temperature, Consistent with a role in initiation, loss of Cdc45p in G(I) prevents all detectable DNA replication without preventing subsequent entry into mitosis, Loss of Cdc45p activity during S-phase blocks S-phase completion but not activation of replication checkpoints. Using density substitution, me show that after allowing replication fork establishment, Cdc45p inactivation prevents the subsequent progression of individual replication forks, This provides the first direct functional evidence that Cdc45p plays an essential role during elongation. Thus, like the large T antigen in SV40 replication, Cdc45p plays a central role in both initiation and elongation phases of chromosomal DNA replication.",
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    author = "JA Tercero and K Labib and JFX Diffley",
    year = "2000",
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    doi = "10.1093/emboj/19.9.2082",
    language = "English",
    volume = "19",
    pages = "2082--2093",
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    DNA synthesis at individual replication forks requires the essential initiation factor Cdc45p. / Tercero, JA; Labib, K; Diffley, JFX.

    In: EMBO Journal, Vol. 19, No. 9, 02.05.2000, p. 2082-2093.

    Research output: Contribution to journalArticle

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    AU - Tercero, JA

    AU - Labib, K

    AU - Diffley, JFX

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    AB - Cdc45p assembles at replication origins before initiation and is required for origin firing in Saccharomyces cerevisiae. A heat-inducible cdc45 degron mutant was constructed that promotes rapid degradation of Cdc45p at the restrictive temperature, Consistent with a role in initiation, loss of Cdc45p in G(I) prevents all detectable DNA replication without preventing subsequent entry into mitosis, Loss of Cdc45p activity during S-phase blocks S-phase completion but not activation of replication checkpoints. Using density substitution, me show that after allowing replication fork establishment, Cdc45p inactivation prevents the subsequent progression of individual replication forks, This provides the first direct functional evidence that Cdc45p plays an essential role during elongation. Thus, like the large T antigen in SV40 replication, Cdc45p plays a central role in both initiation and elongation phases of chromosomal DNA replication.

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    KW - cell cycle

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    KW - BUDDING YEAST

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