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.
Identification of nuclear pre-replication centers poised for DNA synthesis in Xenopus egg extracts: immunolocalization study of replication protein A.
Adachi Y
,
Laemmli UK
.
???displayArticle.abstract???
We demonstrate by immunofluorescence that a 70-kD protein (P70) purified from Xenopus egg extracts is associated with subnuclear foci (about 200) which we propose to be an assembly of DNA pre-replication centers (preRCs). A cDNA encoding this protein reveals that P70 is the Xenopus homologue of replication protein A (RPA also called RF-A). RPA is know to be a cellular, three-subunit single-stranded DNA binding protein, which assists T-antigen in the assembly of the pre-priming complex in the SV40 replication system. The punctated preRCs exist transiently; they form post-mitotically during the period of nuclear membrane breakdown and disappear during ongoing DNA replication. P70 is homogeneously associated with chromatin at the later stages of the S-phase and is displaced from chromatin post replication, so that P70 cannot be detected on mitotic chromosomes. Double-immunofluorescence studies using biotin-dUTP demonstrate that initiation of DNA synthesis is confined to preRCs, resulting in the punctated replication pattern observed previously by others. PreRCs form efficiently on decondensed chromatin in membrane-free egg extracts if ATP and divalent cations are present. Our results suggest that preRCs are composed of an assembly of a large number of pre-initiation replication complexes poised for initiation at discreet subnuclear regions prior to nuclear reconstruction and initiation of DNA synthesis.
Adachi,
Chromosome assembly in vitro: topoisomerase II is required for condensation.
1991, Pubmed,
Xenbase
Adachi,
Chromosome assembly in vitro: topoisomerase II is required for condensation.
1991,
Pubmed
,
Xenbase
Adachi,
Higher order chromosome structure is affected by cold-sensitive mutations in a Schizosaccharomyces pombe gene crm1+ which encodes a 115-kD protein preferentially localized in the nucleus and its periphery.
1989,
Pubmed
Aebersold,
Internal amino acid sequence analysis of proteins separated by one- or two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose.
1987,
Pubmed
Alberts,
Studies with DNA-cellulose chromatography. I. DNA-binding proteins from Escherichia coli.
1968,
Pubmed
Arndt-Jovin,
Covalent attachment of DNA to agarose. Improved synthesis and use in affinity chromatography.
1975,
Pubmed
Berezney,
Nuclear protein matrix: association with newly synthesized DNA.
1975,
Pubmed
Blow,
A role for the nuclear envelope in controlling DNA replication within the cell cycle.
1988,
Pubmed
,
Xenbase
Blow,
A cdc2-like protein is involved in the initiation of DNA replication in Xenopus egg extracts.
1990,
Pubmed
,
Xenbase
Blow,
Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
1986,
Pubmed
,
Xenbase
Bravo,
Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites.
1987,
Pubmed
Brill,
Replication factor-A from Saccharomyces cerevisiae is encoded by three essential genes coordinately expressed at S phase.
1991,
Pubmed
Brill,
Yeast replication factor-A functions in the unwinding of the SV40 origin of DNA replication.
1989,
Pubmed
Challberg,
Animal virus DNA replication.
1989,
Pubmed
Cook,
The nucleoskeleton and the topology of replication.
1991,
Pubmed
Cox,
DNA replication occurs at discrete sites in pseudonuclei assembled from purified DNA in vitro.
1991,
Pubmed
,
Xenbase
de Bruyn Kops,
Formation of DNA replication structures in herpes virus-infected cells requires a viral DNA binding protein.
1988,
Pubmed
Earnshaw,
Topoisomerase II is a structural component of mitotic chromosome scaffolds.
1985,
Pubmed
Erdile,
Characterization of a cDNA encoding the 70-kDa single-stranded DNA-binding subunit of human replication protein A and the role of the protein in DNA replication.
1991,
Pubmed
Fang,
Evidence that the G1-S and G2-M transitions are controlled by different cdc2 proteins in higher eukaryotes.
1991,
Pubmed
,
Xenbase
Finlay,
Reconstitution of biochemically altered nuclear pores: transport can be eliminated and restored.
1990,
Pubmed
,
Xenbase
Fox,
Spatial and temporal distribution of DNA replication sites localized by immunofluorescence and confocal microscopy in mouse fibroblasts.
1991,
Pubmed
Gasser,
Metaphase chromosome structure. Involvement of topoisomerase II.
1986,
Pubmed
Gassmann,
Efficient production of chicken egg yolk antibodies against a conserved mammalian protein.
1990,
Pubmed
Hand,
Eucaryotic DNA: organization of the genome for replication.
1978,
Pubmed
Heyer,
An essential Saccharomyces cerevisiae single-stranded DNA binding protein is homologous to the large subunit of human RP-A.
1990,
Pubmed
Hutchison,
Periodic DNA synthesis in cell-free extracts of Xenopus eggs.
1987,
Pubmed
,
Xenbase
Hutchison,
Changes in the nuclear distribution of DNA polymerase alpha and PCNA/cyclin during the progress of the cell cycle, in a cell-free extract of Xenopus eggs.
1989,
Pubmed
,
Xenbase
Jackson,
Replication occurs at a nucleoskeleton.
1986,
Pubmed
Kenny,
Multiple functions of human single-stranded-DNA binding protein in simian virus 40 DNA replication: single-strand stabilization and stimulation of DNA polymerases alpha and delta.
1989,
Pubmed
Kill,
The timing of the formation and usage of replicase clusters in S-phase nuclei of human diploid fibroblasts.
1991,
Pubmed
,
Xenbase
Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed
Leno,
The nuclear membrane determines the timing of DNA replication in Xenopus egg extracts.
1991,
Pubmed
,
Xenbase
Li,
Simian virus 40 DNA replication in vitro.
1984,
Pubmed
Lohka,
Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts.
1985,
Pubmed
,
Xenbase
Lohka,
Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components.
1983,
Pubmed
,
Xenbase
Lohka,
Roles of cytosol and cytoplasmic particles in nuclear envelope assembly and sperm pronuclear formation in cell-free preparations from amphibian eggs.
1984,
Pubmed
,
Xenbase
Méchali,
DNA synthesis in a cell-free system from Xenopus eggs: priming and elongation on single-stranded DNA in vitro.
1982,
Pubmed
,
Xenbase
Meier,
The role of lamin LIII in nuclear assembly and DNA replication, in cell-free extracts of Xenopus eggs.
1991,
Pubmed
,
Xenbase
Mills,
Replication occurs at discrete foci spaced throughout nuclei replicating in vitro.
1989,
Pubmed
,
Xenbase
Murray,
Cyclin synthesis drives the early embryonic cell cycle.
1989,
Pubmed
,
Xenbase
Murray,
Cell cycle extracts.
1991,
Pubmed
Nakagawa,
A somatic cell-derived system for studying both early and late mitotic events in vitro.
1989,
Pubmed
Nakamura,
Structural organizations of replicon domains during DNA synthetic phase in the mammalian nucleus.
1986,
Pubmed
Nakayasu,
Mapping replicational sites in the eucaryotic cell nucleus.
1989,
Pubmed
Newmeyer,
Assembly in vitro of nuclei active in nuclear protein transport: ATP is required for nucleoplasmin accumulation.
1986,
Pubmed
,
Xenbase
Newport,
Nuclear reconstitution in vitro: stages of assembly around protein-free DNA.
1987,
Pubmed
,
Xenbase
Newport,
Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes.
1987,
Pubmed
,
Xenbase
Newport,
A lamin-independent pathway for nuclear envelope assembly.
1990,
Pubmed
,
Xenbase
Pfaller,
Assembly/disassembly of the nuclear envelope membrane: cell cycle-dependent binding of nuclear membrane vesicles to chromatin in vitro.
1991,
Pubmed
,
Xenbase
Philpott,
Sperm decondensation in Xenopus egg cytoplasm is mediated by nucleoplasmin.
1991,
Pubmed
,
Xenbase
Quinlan,
The intranuclear location of a herpes simplex virus DNA-binding protein is determined by the status of viral DNA replication.
1984,
Pubmed
Rebagliati,
Identification and cloning of localized maternal RNAs from Xenopus eggs.
1985,
Pubmed
,
Xenbase
Ruyechan,
The major herpes simplex virus DNA-binding protein holds single-stranded DNA in an extended configuration.
1983,
Pubmed
Sanger,
DNA sequencing with chain-terminating inhibitors.
1977,
Pubmed
Sheehan,
Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs.
1988,
Pubmed
,
Xenbase
Smith,
Identification, developmental regulation, and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: application of an improved method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots.
1984,
Pubmed
Stillman,
Initiation of eukaryotic DNA replication in vitro.
1989,
Pubmed
Stillman,
Replication and supercoiling of simian virus 40 DNA in cell extracts from human cells.
1985,
Pubmed
Studier,
Use of T7 RNA polymerase to direct expression of cloned genes.
1990,
Pubmed
Towbin,
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
1979,
Pubmed
Tsurimoto,
Sequential initiation of lagging and leading strand synthesis by two different polymerase complexes at the SV40 DNA replication origin.
1990,
Pubmed
Tsurimoto,
Multiple replication factors augment DNA synthesis by the two eukaryotic DNA polymerases, alpha and delta.
1989,
Pubmed
Vaughn,
Replication forks are associated with the nuclear matrix.
1990,
Pubmed
Wobbe,
Replication of simian virus 40 origin-containing DNA in vitro with purified proteins.
1987,
Pubmed
Wobbe,
In vitro replication of duplex circular DNA containing the simian virus 40 DNA origin site.
1985,
Pubmed
Wold,
Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA.
1988,
Pubmed
Wold,
Initiation of simian virus 40 DNA replication in vitro: large-tumor-antigen- and origin-dependent unwinding of the template.
1987,
Pubmed
Wold,
Identification of cellular proteins required for simian virus 40 DNA replication.
1989,
Pubmed
Wu,
Partial purification and characterization of the maturation-promoting factor from eggs of Xenopus laevis.
1980,
Pubmed
,
Xenbase
