Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Mol Cell Biol
1995 Jun 01;156:2942-54. doi: 10.1128/MCB.15.6.2942.
Show Gene links
Show Anatomy links
Site-specific initiation of DNA replication in Xenopus egg extract requires nuclear structure.
Gilbert DM
,
Miyazawa H
,
DePamphilis ML
.
???displayArticle.abstract???
Previous studies have shown that Xenopus egg extract can initiate DNA replication in purified DNA molecules once the DNA is organized into a pseudonucleus. DNA replication under these conditions is independent of DNA sequence and begins at many sites distributed randomly throughout the molecules. In contrast, DNA replication in the chromosomes of cultured animal cells initiates at specific, heritable sites. Here we show that Xenopus egg extract can initiate DNA replication at specific sites in mammalian chromosomes, but only when the DNA is presented in the form of an intact nucleus. Initiation of DNA synthesis in nuclei isolated from G1-phase Chinese hamster ovary cells was distinguished from continuation of DNA synthesis at preformed replication forks in S-phase nuclei by a delay that preceded DNA synthesis, a dependence on soluble Xenopus egg factors, sensitivity to a protein kinase inhibitor, and complete labeling of nascent DNA chains. Initiation sites for DNA replication were mapped downstream of the amplified dihydrofolate reductase gene region by hybridizing newly replicated DNA to unique probes and by hybridizing Okazaki fragments to the two individual strands of unique probes. When G1-phase nuclei were prepared by methods that preserved the integrity of the nuclear membrane, Xenopus egg extract initiated replication specifically at or near the origin of bidirectional replication utilized by hamster cells (dihydrofolate reductase ori-beta). However, when nuclei were prepared by methods that altered nuclear morphology and damaged the nuclear membrane, preference for initiation at ori-beta was significantly reduced or eliminated. Furthermore, site-specific initiation was not observed with bare DNA substrates, and Xenopus eggs or egg extracts replicated prokaryotic DNA or hamster DNA that did not contain a replication origin as efficiently as hamster DNA containing ori-beta. We conclude that initiation sites for DNA replication in mammalian cells are established prior to S phase by some component of nuclear structure and that these sites can be activated by soluble factors in Xenopus eggs.
Adachi,
Study of the cell cycle-dependent assembly of the DNA pre-replication centres in Xenopus egg extracts.
1994, Pubmed,
Xenbase
Adachi,
Study of the cell cycle-dependent assembly of the DNA pre-replication centres in Xenopus egg extracts.
1994,
Pubmed
,
Xenbase
Adam,
Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
1990,
Pubmed
,
Xenbase
Anachkova,
Replication in the amplified dihydrofolate reductase domain in CHO cells may initiate at two distinct sites, one of which is a repetitive sequence element.
1989,
Pubmed
Ariizumi,
Immunoglobulin heavy chain enhancer is located near or in an initiation zone of chromosomal DNA replication.
1993,
Pubmed
Benavente,
Cell type-specific expression of nuclear lamina proteins during development of Xenopus laevis.
1985,
Pubmed
,
Xenbase
Blow,
Replication of purified DNA in Xenopus egg extract is dependent on nuclear assembly.
1990,
Pubmed
,
Xenbase
Blow,
Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
1986,
Pubmed
,
Xenbase
Blow,
Preventing re-replication of DNA in a single cell cycle: evidence for a replication licensing factor.
1993,
Pubmed
,
Xenbase
Blumenthal,
The units of DNA replication in Drosophila melanogaster chromosomes.
1974,
Pubmed
Bozzoni,
Replication of ribosomal DNA in Xenopus laevis.
1981,
Pubmed
,
Xenbase
Buongiorno-Nardelli,
A relationship between replicon size and supercoiled loop domains in the eukaryotic genome.
1982,
Pubmed
,
Xenbase
Burhans,
DNA replication origins in animal cells: a question of context?
1994,
Pubmed
Burhans,
Emetine allows identification of origins of mammalian DNA replication by imbalanced DNA synthesis, not through conservative nucleosome segregation.
1991,
Pubmed
Burhans,
Identification of an origin of bidirectional DNA replication in mammalian chromosomes.
1990,
Pubmed
Caddle,
Intramolecular DNA triplexes, bent DNA and DNA unwinding elements in the initiation region of an amplified dihydrofolate reductase replicon.
1990,
Pubmed
Caddle,
Analysis of the autonomous replication behavior in human cells of the dihydrofolate reductase putative chromosomal origin of replication.
1992,
Pubmed
Carothers,
Structure of the dihydrofolate reductase gene in Chinese hamster ovary cells.
1983,
Pubmed
Carroll,
Localization of a bidirectional DNA replication origin in the native locus and in episomally amplified murine adenosine deaminase loci.
1993,
Pubmed
Church,
Genomic sequencing.
1984,
Pubmed
Coverley,
Regulation of eukaryotic DNA replication.
1994,
Pubmed
Cox,
DNA replication in cell-free extracts from Xenopus eggs is prevented by disrupting nuclear envelope function.
1992,
Pubmed
,
Xenbase
Decker,
Initiation of simian virus 40 DNA replication in vitro: aphidicolin causes accumulation of early-replicating intermediates and allows determination of the initial direction of DNA synthesis.
1986,
Pubmed
DePamphilis,
Eukaryotic DNA replication: anatomy of an origin.
1993,
Pubmed
DePamphilis,
Origins of DNA replication in metazoan chromosomes.
1993,
Pubmed
Diffley,
Two steps in the assembly of complexes at yeast replication origins in vivo.
1994,
Pubmed
,
Xenbase
Dijkwel,
Replication initiation sites are distributed widely in the amplified CHO dihydrofolate reductase domain.
1994,
Pubmed
Dijkwel,
Matrix attachment regions are positioned near replication initiation sites, genes, and an interamplicon junction in the amplified dihydrofolate reductase domain of Chinese hamster ovary cells.
1988,
Pubmed
Dijkwel,
Initiation of DNA replication in the dihydrofolate reductase locus is confined to the early S period in CHO cells synchronized with the plant amino acid mimosine.
1992,
Pubmed
Dinter-Gottlieb,
Aphidicolin arrest irreversibly impairs replicating simian virus 40 chromosomes.
1983,
Pubmed
Dobbs,
Modular sequence elements associated with origin regions in eukaryotic chromosomal DNA.
1994,
Pubmed
Fang,
Distinct roles of cdk2 and cdc2 in RP-A phosphorylation during the cell cycle.
1993,
Pubmed
,
Xenbase
Fox,
Spatial and temporal distribution of DNA replication sites localized by immunofluorescence and confocal microscopy in mouse fibroblasts.
1991,
Pubmed
Gale,
Localization and DNA sequence of a replication origin in the rhodopsin gene locus of Chinese hamster cells.
1992,
Pubmed
Gamper,
Purification of circular DNA using benzoylated naphthoylated DEAE-cellulose.
1985,
Pubmed
Giacca,
Fine mapping of a replication origin of human DNA.
1994,
Pubmed
Gilbert,
Ligand dependence of estrogen receptor induced changes in chromatin structure.
1992,
Pubmed
Gilbert,
Bovine papilloma virus plasmids replicate randomly in mouse fibroblasts throughout S phase of the cell cycle.
1987,
Pubmed
Gilbert,
Site-specific initiation of DNA replication in metazoan chromosomes and the role of nuclear organization.
1993,
Pubmed
,
Xenbase
Gray,
Cell cycle analysis by flow cytometry.
1979,
Pubmed
Haase,
Transcription inhibits the replication of autonomously replicating plasmids in human cells.
1994,
Pubmed
Handeli,
Mapping replication units in animal cells.
1989,
Pubmed
Harland,
Stability of RNA in developing Xenopus embryos and identification of a destabilizing sequence in TFIIIA messenger RNA.
1988,
Pubmed
,
Xenbase
Heintz,
Nuclear DNA synthesis in vitro is mediated via stable replication forks assembled in a temporally specific fashion in vivo.
1988,
Pubmed
Hozák,
Visualization of replication factories attached to nucleoskeleton.
1993,
Pubmed
Hyrien,
Chromosomal replication initiates and terminates at random sequences but at regular intervals in the ribosomal DNA of Xenopus early embryos.
1993,
Pubmed
,
Xenbase
Ishimi,
DNA replication from initiation zones of mammalian cells in a model system.
1994,
Pubmed
Jackson,
Replication occurs at a nucleoskeleton.
1986,
Pubmed
Jenkins,
Nuclei that lack a lamina accumulate karyophilic proteins and assemble a nuclear matrix.
1993,
Pubmed
,
Xenbase
Kill,
The timing of the formation and usage of replicase clusters in S-phase nuclei of human diploid fibroblasts.
1991,
Pubmed
,
Xenbase
Kimelman,
The events of the midblastula transition in Xenopus are regulated by changes in the cell cycle.
1987,
Pubmed
,
Xenbase
Kitsberg,
Replication structure of the human beta-globin gene domain.
1993,
Pubmed
Krysan,
Autonomous replication in human cells of multimers of specific human and bacterial DNA sequences.
1993,
Pubmed
Kubota,
Determination of initiation of DNA replication before and after nuclear formation in Xenopus egg cell free extracts.
1993,
Pubmed
,
Xenbase
Leibovici,
Nuclear distribution of PCNA during embryonic development in Xenopus laevis: a reinvestigation of early cell cycles.
1992,
Pubmed
,
Xenbase
Leno,
The nuclear membrane prevents replication of human G2 nuclei but not G1 nuclei in Xenopus egg extract.
1992,
Pubmed
,
Xenbase
Leu,
High-resolution mapping of replication fork movement through the amplified dihydrofolate reductase domain in CHO cells by in-gel renaturation analysis.
1989,
Pubmed
Liang,
Analysis of an origin of DNA amplification in Sciara coprophila by a novel three-dimensional gel method.
1994,
Pubmed
Lin,
DNA helical instability facilitates initiation at the SV40 replication origin.
1994,
Pubmed
Little,
Initiation and termination of DNA replication in human rRNA genes.
1993,
Pubmed
Mahbubani,
DNA replication initiates at multiple sites on plasmid DNA in Xenopus egg extracts.
1992,
Pubmed
,
Xenbase
Masukata,
Autonomous replication of human chromosomal DNA fragments in human cells.
1993,
Pubmed
McKnight,
Electron microscopic analysis of chromatin replication in the cellular blastoderm Drosophila melanogaster embryo.
1977,
Pubmed
McNeil,
A method for incorporating macromolecules into adherent cells.
1984,
Pubmed
Melera,
Antifolate-resistant Chinese hamster cells. Molecular basis for the biochemical and structural heterogeneity among dihydrofolate reductases produced by drug-sensitive and drug-resistant cell lines.
1988,
Pubmed
Montag,
Structural analysis of the mitotic cycle in pre-gastrula Xenopus embryos.
1988,
Pubmed
,
Xenbase
Nakamura,
Intracellular localization and metabolism of DNA polymerase alpha in human cells visualized with monoclonal antibody.
1984,
Pubmed
Nakayasu,
Mapping replicational sites in the eucaryotic cell nucleus.
1989,
Pubmed
Natale,
Ease of DNA unwinding is a conserved property of yeast replication origins.
1993,
Pubmed
Nethanel,
Two DNA polymerases may be required for synthesis of the lagging DNA strand of simian virus 40.
1990,
Pubmed
Newport,
Nuclear reconstitution in vitro: stages of assembly around protein-free DNA.
1987,
Pubmed
,
Xenbase
Newport,
A lamin-independent pathway for nuclear envelope assembly.
1990,
Pubmed
,
Xenbase
O'Keefe,
Dynamic organization of DNA replication in mammalian cell nuclei: spatially and temporally defined replication of chromosome-specific alpha-satellite DNA sequences.
1992,
Pubmed
Orr-Weaver,
Drosophila chorion genes: cracking the eggshell's secrets.
1991,
Pubmed
Prioleau,
Competition between chromatin and transcription complex assembly regulates gene expression during early development.
1994,
Pubmed
,
Xenbase
Shinomiya,
Analysis of chromosomal replicons in early embryos of Drosophila melanogaster by two-dimensional gel electrophoresis.
1991,
Pubmed
Shinomiya,
Mapping an initiation region of DNA replication at a single-copy chromosomal locus in Drosophila melanogaster cells by two-dimensional gel methods and PCR-mediated nascent-strand analysis: multiple replication origins in a broad zone.
1994,
Pubmed
Sleeman,
Patterns of DNA replication in Drosophila polytene nuclei replicating in Xenopus egg and oocyte extracts.
1992,
Pubmed
,
Xenbase
Snapka,
Aphidicolin-induced topological and recombinational events in simian virus 40.
1991,
Pubmed
Stick,
Changes in the nuclear lamina composition during early development of Xenopus laevis.
1985,
Pubmed
,
Xenbase
Taira,
A novel DNA replication origin identified in the human heat shock protein 70 gene promoter.
1994,
Pubmed
Tasheva,
A mammalian origin of bidirectional DNA replication within the Chinese hamster RPS14 locus.
1994,
Pubmed
Tasheva,
Densely methylated DNA islands in mammalian chromosomal replication origins.
1994,
Pubmed
Trask,
Early dihydrofolate reductase gene amplification events in CHO cells usually occur on the same chromosome arm as the original locus.
1989,
Pubmed
Virta-Pearlman,
Analysis of a replication initiation sequence from the adenosine deaminase region of the mouse genome.
1993,
Pubmed
Wang,
Eukaryotic DNA polymerases.
1991,
Pubmed
Weaver,
The termination region for SV40 DNA replication directs the mode of separation for the two sibling molecules.
1985,
Pubmed
Wolffe,
The role of transcription factors, chromatin structure and DNA replication in 5 S RNA gene regulation.
1994,
Pubmed
,
Xenbase
Wu,
In vivo activity for initiation of DNA replication resides in a transcribed region of the human genome.
1993,
Pubmed
Yamamoto,
Tight association of DNA polymerase alpha with granular structures in the nuclear matrix of chick embryo cell: immunocytochemical detection with monoclonal antibody against DNA polymerase alpha.
1984,
Pubmed
Yoon,
Mapping of replication initiation sites in human ribosomal DNA by nascent-strand abundance analysis.
1995,
Pubmed
