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.
Cell cycle control of higher-order chromatin assembly around naked DNA in vitro.
Hirano T
,
Mitchison TJ
.
???displayArticle.abstract???
We have developed an in vitro system in which higher-order chromatin structures are assembled around naked DNAs in a cell cycle-dependent manner. Membrane-free soluble extracts specific to interphase and mitotic states were prepared from Xenopus eggs. When high molecular weight DNA is incubated with interphase extracts, fluffy chromatin-like structures are assembled. In contrast, mitotic extracts produce highly condensed chromosome-like structures. Immunofluorescence studies show that a monoclonal antibody MPM-2, which recognizes a class of mitosis-specific phosphoproteins, stains the "core" or "axis" of condensed mitotic chromatin but not interphase chromatin. By adding mitotic extracts, interphase chromatin structures are synchronously converted into the condensed state. The increasingly condensed state of chromatin correlates with the appearance and structural rearrangements of the MPM-2-stained structures. These results suggest that mitosis-specific phosphoproteins recognized by MPM-2 may be directly involved in the assembly of the chromosome scaffold-like structures and chromatin condensation. Although both extracts promote nucleosome assembly at the same rate, topoisomerase II (topo II) activity is four to five times higher in mitotic extracts compared with interphase extracts. The addition of a topo II inhibitor VM-26 into mitotic assembly mixtures disturbs the organization of the MPM-2-stained structures and affects the final stage of chromatin condensation. This in vitro system should be useful for identifying cis- and trans-acting elements responsible for higher-order chromatin assembly and its structural changes in the cell cycle.
Ackerman,
Phosphorylation of DNA topoisomerase II in vivo and in total homogenates of Drosophila Kc cells. The role of casein kinase II.
1988, Pubmed
Ackerman,
Phosphorylation of DNA topoisomerase II in vivo and in total homogenates of Drosophila Kc cells. The role of casein kinase II.
1988,
Pubmed
Ackerman,
Phosphorylation of DNA topoisomerase II by casein kinase II: modulation of eukaryotic topoisomerase II activity in vitro.
1985,
Pubmed
Adachi,
Chromosome assembly in vitro: topoisomerase II is required for condensation.
1991,
Pubmed
,
Xenbase
Allan,
Cell cycle control of microtubule-based membrane transport and tubule formation in vitro.
1991,
Pubmed
,
Xenbase
Amati,
Chromosomal ARS and CEN elements bind specifically to the yeast nuclear scaffold.
1988,
Pubmed
Belmont,
Real-time visualization of cell cycle-dependent changes in microtubule dynamics in cytoplasmic extracts.
1990,
Pubmed
,
Xenbase
Benavente,
Cell type-specific expression of nuclear lamina proteins during development of Xenopus laevis.
1985,
Pubmed
,
Xenbase
Blow,
Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs.
1986,
Pubmed
,
Xenbase
Blow,
Replication of purified DNA in Xenopus egg extract is dependent on nuclear assembly.
1990,
Pubmed
,
Xenbase
Charron,
DNA topoisomerase II is required for formation of mitotic chromosomes in Chinese hamster ovary cells: studies using the inhibitor 4'-demethylepipodophyllotoxin 9-(4,6-O-thenylidene-beta-D-glucopyranoside).
1990,
Pubmed
Chen,
Nonintercalative antitumor drugs interfere with the breakage-reunion reaction of mammalian DNA topoisomerase II.
1984,
Pubmed
Cockerill,
Chromosomal loop anchorage of the kappa immunoglobulin gene occurs next to the enhancer in a region containing topoisomerase II sites.
1986,
Pubmed
Davis,
Monoclonal antibodies to mitotic cells.
1983,
Pubmed
,
Xenbase
Dilworth,
Two complexes that contain histones are required for nucleosome assembly in vitro: role of nucleoplasmin and N1 in Xenopus egg extracts.
1987,
Pubmed
,
Xenbase
Earnshaw,
Topoisomerase II is a structural component of mitotic chromosome scaffolds.
1985,
Pubmed
Estey,
Cell cycle stage dependent variations in drug-induced topoisomerase II mediated DNA cleavage and cytotoxicity.
1987,
Pubmed
Gasser,
Metaphase chromosome structure. Involvement of topoisomerase II.
1986,
Pubmed
Gasser,
Studies on scaffold attachment sites and their relation to genome function.
1989,
Pubmed
Gurley,
Histone phosphorylation and chromatin structure during mitosis in Chinese hamster cells.
1978,
Pubmed
Heck,
In vivo phosphorylation of the 170-kDa form of eukaryotic DNA topoisomerase II. Cell cycle analysis.
1989,
Pubmed
Hirano,
A temperature-sensitive mutation of the Schizosaccharomyces pombe gene nuc2+ that encodes a nuclear scaffold-like protein blocks spindle elongation in mitotic anaphase.
1988,
Pubmed
Hirano,
Snap helix with knob and hole: essential repeats in S. pombe nuclear protein nuc2+.
1990,
Pubmed
Hofmann,
RAP-1 factor is necessary for DNA loop formation in vitro at the silent mating type locus HML.
1989,
Pubmed
Kayne,
Extremely conserved histone H4 N terminus is dispensable for growth but essential for repressing the silent mating loci in yeast.
1988,
Pubmed
Laskey,
Assembly of SV40 chromatin in a cell-free system from Xenopus eggs.
1977,
Pubmed
,
Xenbase
Laskey,
Assembly of the cell nucleus.
1990,
Pubmed
Lohka,
Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts.
1985,
Pubmed
,
Xenbase
Matthews,
The role of H1 histone phosphorylation in the cell cycle. Turbidity studies of H1-DNA interaction.
1978,
Pubmed
Mirkovitch,
Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold.
1984,
Pubmed
Mitchison,
Properties of the kinetochore in vitro. I. Microtubule nucleation and tubulin binding.
1985,
Pubmed
Moreno,
Substrates for p34cdc2: in vivo veritas?
1990,
Pubmed
Murray,
The role of cyclin synthesis and degradation in the control of maturation promoting factor activity.
1989,
Pubmed
,
Xenbase
Murray,
Cyclin synthesis drives the early embryonic cell cycle.
1989,
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
Paulson,
The structure of histone-depleted metaphase chromosomes.
1977,
Pubmed
Roberge,
The topoisomerase II inhibitor VM-26 induces marked changes in histone H1 kinase activity, histones H1 and H3 phosphorylation, and chromosome condensation in G2 phase and mitotic BHK cells.
1990,
Pubmed
Sahyoun,
Protein kinase C phosphorylates topoisomerase II: topoisomerase activation and its possible role in phorbol ester-induced differentiation of HL-60 cells.
1986,
Pubmed
Saijo,
Purification and characterization of type II DNA topoisomerase from mouse FM3A cells: phosphorylation of topoisomerase II and modification of its activity.
1990,
Pubmed
Sawin,
Mitotic spindle assembly by two different pathways in vitro.
1991,
Pubmed
,
Xenbase
Thoma,
Involvement of histone H1 in the organization of the nucleosome and of the salt-dependent superstructures of chromatin.
1979,
Pubmed
Uemura,
DNA topoisomerase II is required for condensation and separation of mitotic chromosomes in S. pombe.
1987,
Pubmed
Vale,
Severing of stable microtubules by a mitotically activated protein in Xenopus egg extracts.
1991,
Pubmed
,
Xenbase
Vandre,
Phosphoproteins are components of mitotic microtubule organizing centers.
1984,
Pubmed
Vandré,
Distribution of cytoskeletal proteins sharing a conserved phosphorylated epitope.
1986,
Pubmed
Verde,
Regulation of microtubule dynamics by cdc2 protein kinase in cell-free extracts of Xenopus eggs.
1990,
Pubmed
,
Xenbase
Wood,
Mitotic chromatin condensation in vitro using somatic cell extracts and nuclei with variable levels of endogenous topoisomerase II.
1990,
Pubmed
Wright,
Teniposide, a topoisomerase II inhibitor, prevents chromosome condensation and separation but not decondensation in fertilized surf clam (Spisula solidissima) oocytes.
1990,
Pubmed
