Replication factory activation can be decoupled from the replication timing program by modulating Cdk levels

Alexander M. Thomson, Peter J. Gillespie, J. Julian Blow (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    41 Citations (Scopus)
    144 Downloads (Pure)

    Abstract

    In the metazoan replication timing program, clusters of replication origins located in different subchromosomal domains fire at different times during S phase. We have used Xenopus laevis egg extracts to drive an accelerated replication timing program in mammalian nuclei. Although replicative stress caused checkpoint-induced slowing of the timing program, inhibition of checkpoint kinases in an unperturbed S phase did not accelerate it. Lowering cyclin-dependent kinase (Cdk) activity slowed both replication rate and progression through the timing program, whereas raising Cdk activity increased them. Surprisingly, modest alteration of Cdk activity changed the amount of DNA synthesized during different stages of the timing program. This was associated with a change in the number of active replication factories, whereas the distribution of origins within active factories remained relatively normal. The ability of Cdks to differentially effect replication initiation, factory activation, and progression through the timing program provides new insights into the way that chromosomal DNA replication is organized during S phase.

    Original languageEnglish
    Pages (from-to)209-221
    Number of pages13
    JournalJournal of Cell Biology
    Volume188
    Issue number2
    DOIs
    Publication statusPublished - 25 Jan 2010

    Keywords

    • CYCLIN-DEPENDENT KINASES
    • XENOPUS EGG EXTRACTS
    • S-PHASE CHECKPOINT
    • DNA-REPLICATION
    • CELL-CYCLE
    • DORMANT ORIGINS
    • BUDDING YEAST
    • FISSION YEAST
    • CHROMOSOMAL DOMAINS
    • REPLICON CLUSTERS

    Cite this

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    title = "Replication factory activation can be decoupled from the replication timing program by modulating Cdk levels",
    abstract = "In the metazoan replication timing program, clusters of replication origins located in different subchromosomal domains fire at different times during S phase. We have used Xenopus laevis egg extracts to drive an accelerated replication timing program in mammalian nuclei. Although replicative stress caused checkpoint-induced slowing of the timing program, inhibition of checkpoint kinases in an unperturbed S phase did not accelerate it. Lowering cyclin-dependent kinase (Cdk) activity slowed both replication rate and progression through the timing program, whereas raising Cdk activity increased them. Surprisingly, modest alteration of Cdk activity changed the amount of DNA synthesized during different stages of the timing program. This was associated with a change in the number of active replication factories, whereas the distribution of origins within active factories remained relatively normal. The ability of Cdks to differentially effect replication initiation, factory activation, and progression through the timing program provides new insights into the way that chromosomal DNA replication is organized during S phase.",
    keywords = "CYCLIN-DEPENDENT KINASES, XENOPUS EGG EXTRACTS, S-PHASE CHECKPOINT, DNA-REPLICATION, CELL-CYCLE, DORMANT ORIGINS, BUDDING YEAST, FISSION YEAST, CHROMOSOMAL DOMAINS, REPLICON CLUSTERS",
    author = "Thomson, {Alexander M.} and Gillespie, {Peter J.} and Blow, {J. Julian}",
    year = "2010",
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    doi = "10.1083/jcb.200911037",
    language = "English",
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    pages = "209--221",
    journal = "Journal of Cell Biology",
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    TY - JOUR

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    AU - Gillespie, Peter J.

    AU - Blow, J. Julian

    PY - 2010/1/25

    Y1 - 2010/1/25

    N2 - In the metazoan replication timing program, clusters of replication origins located in different subchromosomal domains fire at different times during S phase. We have used Xenopus laevis egg extracts to drive an accelerated replication timing program in mammalian nuclei. Although replicative stress caused checkpoint-induced slowing of the timing program, inhibition of checkpoint kinases in an unperturbed S phase did not accelerate it. Lowering cyclin-dependent kinase (Cdk) activity slowed both replication rate and progression through the timing program, whereas raising Cdk activity increased them. Surprisingly, modest alteration of Cdk activity changed the amount of DNA synthesized during different stages of the timing program. This was associated with a change in the number of active replication factories, whereas the distribution of origins within active factories remained relatively normal. The ability of Cdks to differentially effect replication initiation, factory activation, and progression through the timing program provides new insights into the way that chromosomal DNA replication is organized during S phase.

    AB - In the metazoan replication timing program, clusters of replication origins located in different subchromosomal domains fire at different times during S phase. We have used Xenopus laevis egg extracts to drive an accelerated replication timing program in mammalian nuclei. Although replicative stress caused checkpoint-induced slowing of the timing program, inhibition of checkpoint kinases in an unperturbed S phase did not accelerate it. Lowering cyclin-dependent kinase (Cdk) activity slowed both replication rate and progression through the timing program, whereas raising Cdk activity increased them. Surprisingly, modest alteration of Cdk activity changed the amount of DNA synthesized during different stages of the timing program. This was associated with a change in the number of active replication factories, whereas the distribution of origins within active factories remained relatively normal. The ability of Cdks to differentially effect replication initiation, factory activation, and progression through the timing program provides new insights into the way that chromosomal DNA replication is organized during S phase.

    KW - CYCLIN-DEPENDENT KINASES

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    KW - S-PHASE CHECKPOINT

    KW - DNA-REPLICATION

    KW - CELL-CYCLE

    KW - DORMANT ORIGINS

    KW - BUDDING YEAST

    KW - FISSION YEAST

    KW - CHROMOSOMAL DOMAINS

    KW - REPLICON CLUSTERS

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    JO - Journal of Cell Biology

    JF - Journal of Cell Biology

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    ER -