Purification and mechanism of action of a nucleosome assembly factor from Xenopus oocytes.

Chromatin with nucleosomes spaced at 180-base pair intervals can be formed in vitro on circular DNA molecules using a Xenopus oocyte S-150 extract, but the ability to form a periodic chromatin structure is lost upon fractionation of this extract. To identify factors other than the known ones involved in chromatin assembly, we have first depleted the extract by incubating it in batch with charged resins, and we have subsequently reconstituted it with purified fractions. Studies performed with the fractionated components indicate that formation of periodically spaced nucleosomes on the relaxed, closed circular DNA proceeds in two steps and does not require DNA topoisomerases. In a first step, histones H3/H4 are transferred from the endogenous H3/H4-N1 complex to the DNA, forming a nascent chromatin structure. This structure can then be rapidly complemented in a subsequent and independent step with a stoichiometric amount of histone H2A/H2B dimers. Under these experimental conditions, excess histone H2A/H2B dimers inhibit DNA supercoiling and nucleosome formation. We describe the purification of a factor from the Xenopus oocyte S-150 which permits DNA supercoiling and nucleosome formation under conditions of excess histone H2A/H2B. The activity purifies as a complex of five nonacidic polypeptides with apparent molecular masses ranging between 56 and 62 kDa. This factor prevents the binding of excess histone H2A/H2B to the DNA, and it can also remove excess histone H2A/H2B already bound to the DNA, thus ensuring that stoichiometric amounts of all four nucleosomal histones associate with the DNA.

• Publication year

  1990

• Venue

  Journal of Biological Chemistry

• Publication date

  1990-06-05

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1016/s0021-9258(19)38857-xPMID 2160975
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• The transcription complex of the 5 S RNA gene, but not transcription factor IIIA alone, prevents nucleosomal repression of transcription.

  1990cited by this paper
• Purification and characterization of CAF-I, a human cell factor required for chromatin assembly during DNA replication in vitro.

  1989cited by this paper
• Assembly and properties of chromatin containing histone H1.

  1989cited by this paper
• The assembly of regularly spaced nucleosomes in the Xenopus oocyte S-150 extract is accompanied by deacetylation of histone H4.

  1989cited by this paper
• Expression of a histone H1-like protein is restricted to early Xenopus development.

  1988cited by this paper
• Characterization of the repressed 5S DNA minichromosomes assembled in vitro with a high-speed supernatant of Xenopus laevis oocytes

  1988cited by this paper
• Chromatin assembly in vitro and in vivo

  1988cited by this paper
• Assembly of spaced chromatin involvement of ATP and DNA topoisomerase activity.

  1988cited by this paper
• Monoclonal antibodies neutralizing mammalian DNA topoisomerase I activity.

  1988cited by this paper
• Two complexes that contain histones are required for nucleosome assembly in vitro: role of nucleoplasmin and N1 in Xenopus egg extracts.

  1987cited by this paper
• Chromatin reconstituted from tandemly repeated cloned DNA fragments and core histones: a model system for study of higher order structure.

  1985cited by this paper
• Co-existence of two different types of soluble histone complexes in nuclei of Xenopus laevis oocytes.

  1985cited by this paper
• Chromatin assembly in Xenopus oocytes: in vitro studies.

  1984cited by this paper
• Simian virus 40 DNA replication in vitro.

  1984cited by this paper
• Purification and initial characterization of a protein which facilitates assembly of nucleosome-like structure from mammalian cells.

  1984cited by this paper
• A protein which facilitates assembly of nucleosome-like structures in vitro in mammalian cells.

  1983cited by this paper
• A simple procedure for parallel sequence analysis of both strands of 5'-labeled DNA.

  1983cited by this paper
• Association of newly synthesized histones with replicating and nonreplicating regions of chromatin.

  1982cited by this paper
• Soluble acidic complexes containing histones H3 and H4 in nuclei of Xenopus laevis oocytes.

  1982cited by this paper
• A new method for the isolation of replicative chromatin: selective deposition of histone on both new and old DNA.

  1981cited by this paper
• Nucleosome assembly

  1980cited by this paper
• Sequential assembly of newly synthesized histones on replicating SV40 DNA.

  1980cited by this paper
• Assembly of nucleosomes: the reaction involving X. laevis nucleoplasmin.

  1980cited by this paper
• Chromatin structure of the 5S ribonucleic acid genes of Xenopus laevis.

  1979cited by this paper
• Assembly of newly replicated chromatin.

  1978cited by this paper
• Selective DNA conservation and chromatin assembly after injection of SV40 DNA into Xenopus oocytes.

  1978cited by this paper
• Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA

  1978cited by this paper
• Preferential association of newly synthesized H3 and H4 histones with newly replicated DNA.

  1978cited by this paper
• Assembly of SV40 chromatin in a cell-free system from Xenopus eggs.

  1977cited by this paper
• Supercoiling energy and nucleosome formation: the role of the arginine-rich histone kernel.

  1977cited by this paper
• The organization of histones and DNA in chromatin: evidence for an arginine-rich histone kernel.

  1976cited by this paper
• Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

  1970cited by this paper

• Histone Octamer Dissociation Is Not Required for in Vitro Replication of Simian Virus 40 Minichromosomes*

  2000cites this paper
• Purification and Mechanism of Action of a Nucleosome Assembly Factor from Xenopus Oocytes*

  2000cites this paper
• Characterization of the Histone H1-binding Protein, NASP, as a Cell Cycle-regulated Somatic Protein*

  2000cites this paper
• Variable Structure and Folding of DNA

  1998cites this paper
• Structural and functional analysis of chromatin assembled from defined histones.

  1998cites this paper
• Nuclear reassemblyin vitro is independent of nucleosome/chromatin assembly

  1998cites this paper
• Histones in transit: cytosolic histone complexes and diacetylation of H4 during nucleosome assembly in human cells.

  1997cites this paper
• Identification of a small, very acidic constitutive nucleolar protein (NO29) as a member of the nucleoplasmin family.

  1997cites this paper
• Topoisomerase function during replication-independent chromatin assembly in yeast

  1997cites this paper
• Dissociation of protamine-DNA complexes by Xenopus nucleoplasmin and minichromosome assembly in vitro.

  1996cites this paper
• Drosophila NAP-1 is a core histone chaperone that functions in ATP-facilitated assembly of regularly spaced nucleosomal arrays

  1996cites this paper
• Structure of replicating chromatin.

  1996cites this paper
• Assembly of regularly spaced nucleosome arrays by Drosophila chromatin assembly factor 1 and a 56-kDa histone-binding protein.

  1995cites this paper
• Molecular cloning and functional characterization of a cDNA encoding nucleosome assembly protein 1 (NAP-1) from soybean

  1995cites this paper
• Histone Acetylation During Chromatin Replication and Nucleosome Assembly

  1995cites this paper
• A histone‐binding protein, nucleoplasmin, stimulates transcription factor binding to nucleosomes and factor‐induced nucleosome disassembly.

  1994cites this paper
• Analysis of nucleosome assembly and histone exchange using antibodies specific for acetylated H4.

  1993cites this paper
• Disruption of the nucleosomes at the replication fork.

  1993cites this paper
• Nuclear assembly, structure, and function: the use of Xenopus in vitro systems.

  1993cites this paper
• The role of nucleoplasmin in chromatin assembly and disassembly.

  1993cites this paper
• Partial purification, from Xenopus laevis oocytes, of an ATP-dependent activity required for nucleosome spacing in vitro.

  1992cites this paper
• Cell-free system for assembly of transcriptionally repressed chromatin from Drosophila embryos.

  1992cites this paper
• Cell-free system for assembly of transcriptionally repressed chromatin from Drosophila embryos

  1992cites this paper
• Chromatin replication

  1992cites this paper
• Nucleosome core displacement in vitro via a metastable transcription factor-nucleosome complex.

  1992cites this paper
• Comparison of replicative and non-replicative chromatin assembly pathways in HeLa cell extracts.

  1992cites this paper
• Emetine allows identification of origins of mammalian DNA replication by imbalanced DNA synthesis, not through conservative nucleosome segregation.

  1991cites this paper
• DNA‐dependent phosphorylation of histone H2A.X during nucleosome assembly in Xenopus laevis oocytes: involvement of protein phosphorylation in nucleosome spacing.

  1991cites this paper
• Identification and molecular cloning of yeast homolog of nucleosome assembly protein I which facilitates nucleosome assembly in vitro.

  1991cites this paper
• Stepwise assembly of chromatin during DNA replication in vitro.

  1991cites this paper
• Assembly of the cell nucleus.

  1990cites this paper
• The histone H3/H4.N1 complex supplemented with histone H2A-H2B dimers and DNA topoisomerase I forms nucleosomes on circular DNA under physiological conditions.

  1990cites this paper
• Cell-Free System for Assembly of Transcriptionally Repressed Chromatin from Drosophila Embryos

  year unknowncites this paper