Chromosome replication in cell-free systems from Xenopus eggs

J. J. Blow, S. M. Dilworth, C. Dingwall, A. D. Mills, R. A. Laskey

    Research output: Contribution to journalArticle

    25 Citations (Scopus)

    Abstract

    Cell-free systems from eggs of the frog Xenopus laevis are able to perform most of the acts of eukaryotic chromosome replication in vitro. This now includes the crucial regulatory step of initiation, which had only been achieved for viral systems previously. Purified DNA or nuclei are able to initiate and complete semi-conservation replication in egg extracts in vitro (Blow & Laskey, Cell 47, 557-587 (1986). Replication does not require specialized DNA sequences either in vitro or in microinjected eggs, but in both systems large templates replicate more efficiently than small templates. In some cases replication can re-initiate, excluding the possibility that replication is primed by preexisting primers in the template preparations. When nuclei are replicated in vitro, only one round of replication is observed in a single incubation resembling the single round of replication observed for purified DNA after micro-injection. The mechanism that prevents re-initiation of replication within a single cell cycle is discussed and certain models are eliminated. Nucleosome assembly from histones and DNA has also been studied in cell-free systems from Xenopus eggs. Fractionation has led to the identification of two acidic proteins called nucleoplasmin and N1, which bind histones and transfer them to DNA. The sequences of both proteins have been determined by cDNA cloning and sequencing. Both proteins are found as complexes with histones in eggs.
    Original languageEnglish
    Pages (from-to)483-494
    Number of pages12
    JournalPhilosophical Transactions of the Royal Society B - Biological Sciences
    Volume317
    Issue number1187
    Publication statusPublished - 15 Dec 1987

    Fingerprint

    cell free system
    Cell-Free System
    Xenopus
    Chromosomes
    Eggs
    histones
    Histones
    chromosomes
    DNA
    Nucleoplasmins
    Proteins
    nucleosomes
    Nucleosomes
    Cloning
    DNA sequences
    Xenopus laevis
    Fractionation
    Anura
    Ovum
    Organism Cloning

    Cite this

    Blow, J. J. ; Dilworth, S. M. ; Dingwall, C. ; Mills, A. D. ; Laskey, R. A. / Chromosome replication in cell-free systems from Xenopus eggs. In: Philosophical Transactions of the Royal Society B - Biological Sciences. 1987 ; Vol. 317, No. 1187. pp. 483-494.
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    abstract = "Cell-free systems from eggs of the frog Xenopus laevis are able to perform most of the acts of eukaryotic chromosome replication in vitro. This now includes the crucial regulatory step of initiation, which had only been achieved for viral systems previously. Purified DNA or nuclei are able to initiate and complete semi-conservation replication in egg extracts in vitro (Blow & Laskey, Cell 47, 557-587 (1986). Replication does not require specialized DNA sequences either in vitro or in microinjected eggs, but in both systems large templates replicate more efficiently than small templates. In some cases replication can re-initiate, excluding the possibility that replication is primed by preexisting primers in the template preparations. When nuclei are replicated in vitro, only one round of replication is observed in a single incubation resembling the single round of replication observed for purified DNA after micro-injection. The mechanism that prevents re-initiation of replication within a single cell cycle is discussed and certain models are eliminated. Nucleosome assembly from histones and DNA has also been studied in cell-free systems from Xenopus eggs. Fractionation has led to the identification of two acidic proteins called nucleoplasmin and N1, which bind histones and transfer them to DNA. The sequences of both proteins have been determined by cDNA cloning and sequencing. Both proteins are found as complexes with histones in eggs.",
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    Blow, JJ, Dilworth, SM, Dingwall, C, Mills, AD & Laskey, RA 1987, 'Chromosome replication in cell-free systems from Xenopus eggs', Philosophical Transactions of the Royal Society B - Biological Sciences, vol. 317, no. 1187, pp. 483-494.

    Chromosome replication in cell-free systems from Xenopus eggs. / Blow, J. J.; Dilworth, S. M.; Dingwall, C.; Mills, A. D.; Laskey, R. A.

    In: Philosophical Transactions of the Royal Society B - Biological Sciences, Vol. 317, No. 1187, 15.12.1987, p. 483-494.

    Research output: Contribution to journalArticle

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    AU - Blow, J. J.

    AU - Dilworth, S. M.

    AU - Dingwall, C.

    AU - Mills, A. D.

    AU - Laskey, R. A.

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    Y1 - 1987/12/15

    N2 - Cell-free systems from eggs of the frog Xenopus laevis are able to perform most of the acts of eukaryotic chromosome replication in vitro. This now includes the crucial regulatory step of initiation, which had only been achieved for viral systems previously. Purified DNA or nuclei are able to initiate and complete semi-conservation replication in egg extracts in vitro (Blow & Laskey, Cell 47, 557-587 (1986). Replication does not require specialized DNA sequences either in vitro or in microinjected eggs, but in both systems large templates replicate more efficiently than small templates. In some cases replication can re-initiate, excluding the possibility that replication is primed by preexisting primers in the template preparations. When nuclei are replicated in vitro, only one round of replication is observed in a single incubation resembling the single round of replication observed for purified DNA after micro-injection. The mechanism that prevents re-initiation of replication within a single cell cycle is discussed and certain models are eliminated. Nucleosome assembly from histones and DNA has also been studied in cell-free systems from Xenopus eggs. Fractionation has led to the identification of two acidic proteins called nucleoplasmin and N1, which bind histones and transfer them to DNA. The sequences of both proteins have been determined by cDNA cloning and sequencing. Both proteins are found as complexes with histones in eggs.

    AB - Cell-free systems from eggs of the frog Xenopus laevis are able to perform most of the acts of eukaryotic chromosome replication in vitro. This now includes the crucial regulatory step of initiation, which had only been achieved for viral systems previously. Purified DNA or nuclei are able to initiate and complete semi-conservation replication in egg extracts in vitro (Blow & Laskey, Cell 47, 557-587 (1986). Replication does not require specialized DNA sequences either in vitro or in microinjected eggs, but in both systems large templates replicate more efficiently than small templates. In some cases replication can re-initiate, excluding the possibility that replication is primed by preexisting primers in the template preparations. When nuclei are replicated in vitro, only one round of replication is observed in a single incubation resembling the single round of replication observed for purified DNA after micro-injection. The mechanism that prevents re-initiation of replication within a single cell cycle is discussed and certain models are eliminated. Nucleosome assembly from histones and DNA has also been studied in cell-free systems from Xenopus eggs. Fractionation has led to the identification of two acidic proteins called nucleoplasmin and N1, which bind histones and transfer them to DNA. The sequences of both proteins have been determined by cDNA cloning and sequencing. Both proteins are found as complexes with histones in eggs.

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