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.
Study of the cell cycle-dependent assembly of the DNA pre-replication centres in Xenopus egg extracts.
Adachi Y, Laemmli UK.
???displayArticle.abstract??? RPA is a cellular, three-subunit, single-stranded (ss) DNA binding protein, which assists T-antigen in the assembly of the pre-priming complex in the SV40 replication system. By immunodepletion and complementation, we have identified RPA as an essential factor for cellular DNA replication in Xenopus extracts. RPA assembles post-mitotically on the decondensing chromosomes into numerous subnuclear pre-replication centres (preRCs) which serve, upon formation of the nuclear membrane, as RCs for the initiation of DNA synthesis. By a variety of experiments including the use of isolated components, we demonstrate that an inactive cdc2-cyclin B kinase complex is essential to allow post-mitotic assembly of the preRCs. In contrast, the active cdk2-cyclin A kinase does not impede or facilitate the assembly of preRCs. Digestion analysis using the single-strand-specific P1 nuclease as well as competition experiments with ssDNA, reveal that replication-associated unwinding of the DNA, assisted by RPA, requires the formation of the nuclear membrane. The p21 cdk-interacting protein Cip1 appears to inhibit DNA replication prior to the unwinding DNA step, but after assembly of preRC and nuclear reconstruction.
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,
Identification of nuclear pre-replication centers poised for DNA synthesis in Xenopus egg extracts: immunolocalization study of replication protein A.
1992,
Pubmed
,
Xenbase Alberts,
Studies with DNA-cellulose chromatography. I. DNA-binding proteins from Escherichia coli.
1968,
Pubmed Almouzni,
Nuclear assembly, structure, and function: the use of Xenopus in vitro systems.
1993,
Pubmed
,
Xenbase Blow,
Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
1986,
Pubmed
,
Xenbase Blow,
A cdc2-like protein is involved in the initiation of DNA replication in Xenopus egg extracts.
1990,
Pubmed
,
Xenbase Blow,
A role for the nuclear envelope in controlling DNA replication within the cell cycle.
1988,
Pubmed
,
Xenbase Bradford,
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
1976,
Pubmed Bramhill,
Duplex opening by dnaA protein at novel sequences in initiation of replication at the origin of the E. coli chromosome.
1988,
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 Callan,
Replication of DNA in the chromosomes of eukaryotes.
1972,
Pubmed Cardoso,
Reversal of terminal differentiation and control of DNA replication: cyclin A and Cdk2 specifically localize at subnuclear sites of DNA replication.
1993,
Pubmed Carle,
Electrophoretic separations of large DNA molecules by periodic inversion of the electric field.
1986,
Pubmed Challberg,
Animal virus DNA replication.
1989,
Pubmed Coverley,
Requirement for the replication protein SSB in human DNA excision repair.
1991,
Pubmed Cox,
DNA replication in cell-free extracts from Xenopus eggs is prevented by disrupting nuclear envelope function.
1992,
Pubmed
,
Xenbase Desai,
Activation of human cyclin-dependent kinases in vitro.
1992,
Pubmed Din,
Cell-cycle-regulated phosphorylation of DNA replication factor A from human and yeast cells.
1990,
Pubmed Dornreiter,
Interaction of DNA polymerase alpha-primase with cellular replication protein A and SV40 T antigen.
1992,
Pubmed Dunphy,
The Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis.
1988,
Pubmed
,
Xenbase Dutta,
Inhibition of DNA replication factor RPA by p53.
1993,
Pubmed Dutta,
cdc2 family kinases phosphorylate a human cell DNA replication factor, RPA, and activate DNA replication.
1992,
Pubmed Fairman,
Cellular factors required for multiple stages of SV40 DNA replication in vitro.
1988,
Pubmed Fang,
Distinct roles of cdk2 and cdc2 in RP-A phosphorylation during the cell cycle.
1993,
Pubmed
,
Xenbase Fang,
Evidence that the G1-S and G2-M transitions are controlled by different cdc2 proteins in higher eukaryotes.
1991,
Pubmed
,
Xenbase Felix,
A post-ribosomal supernatant from activated Xenopus eggs that displays post-translationally regulated oscillation of its cdc2+ mitotic kinase activity.
1989,
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 Grieco,
An improved procedure for the purification of protein fused with glutathione S-transferase.
1992,
Pubmed Gu,
Inhibition of CDK2 activity in vivo by an associated 20K regulatory subunit.
1993,
Pubmed Harper,
The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases.
1993,
Pubmed He,
The transactivator proteins VP16 and GAL4 bind replication factor A.
1993,
Pubmed Heyer,
An essential Saccharomyces cerevisiae single-stranded DNA binding protein is homologous to the large subunit of human RP-A.
1990,
Pubmed Hovanessian,
Efficiency of poly(A).poly(U) as an adjuvant.
1988,
Pubmed Hunter,
Braking the cycle.
1993,
Pubmed Hutchison,
Periodic DNA synthesis in cell-free extracts of Xenopus eggs.
1987,
Pubmed
,
Xenbase Hyrien,
Chromosomal replication initiates and terminates at random sequences but at regular intervals in the ribosomal DNA of Xenopus early embryos.
1993,
Pubmed
,
Xenbase Jenkins,
Nuclei that lack a lamina accumulate karyophilic proteins and assemble a nuclear matrix.
1993,
Pubmed
,
Xenbase Käs,
In vivo topoisomerase II cleavage of the Drosophila histone and satellite III repeats: DNA sequence and structural characteristics.
1992,
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 Kenny,
The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication.
1990,
Pubmed Kim,
Binding properties of replication protein A from human and yeast cells.
1992,
Pubmed Labbé,
M phase-specific cdc2 kinase: preparation from starfish oocytes and properties.
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 Leno,
The nuclear membrane prevents replication of human G2 nuclei but not G1 nuclei in Xenopus egg extract.
1992,
Pubmed
,
Xenbase Li,
The acidic transcriptional activation domains of VP16 and p53 bind the cellular replication protein A and stimulate in vitro BPV-1 DNA replication.
1993,
Pubmed 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 Lohka,
Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components.
1983,
Pubmed
,
Xenbase Lohka,
Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts.
1985,
Pubmed
,
Xenbase Mahbubani,
DNA replication initiates at multiple sites on plasmid DNA in Xenopus egg extracts.
1992,
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 Miake-Lye,
Maturation-promoting factor induces nuclear envelope breakdown in cycloheximide-arrested embryos of Xenopus laevis.
1983,
Pubmed
,
Xenbase Mills,
Replication occurs at discrete foci spaced throughout nuclei replicating in vitro.
1989,
Pubmed
,
Xenbase Murray,
Cell cycle extracts.
1991,
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,
In vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins.
1986,
Pubmed
,
Xenbase Newport,
A lamin-independent pathway for nuclear envelope assembly.
1990,
Pubmed
,
Xenbase Newport,
Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes.
1987,
Pubmed
,
Xenbase Newport,
Nuclear reconstitution in vitro: stages of assembly around protein-free DNA.
1987,
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 Schnos,
Initiation protein induced helix destabilization at the lambda origin: a prepriming step in DNA replication.
1988,
Pubmed Sheehan,
Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs.
1988,
Pubmed
,
Xenbase Stillman,
Initiation of eukaryotic DNA replication in vitro.
1989,
Pubmed Towbin,
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
1979,
Pubmed Wilcock,
Localization of p53, retinoblastoma and host replication proteins at sites of viral replication in herpes-infected cells.
1991,
Pubmed Wold,
Identification of cellular proteins required for simian virus 40 DNA replication.
1989,
Pubmed Wold,
Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA.
1988,
Pubmed Xiong,
p21 is a universal inhibitor of cyclin kinases.
1993,
Pubmed
