Abstract
Nucleosome-like structures have been efficiently assembled in vitro by interaction of cauliflower histones, pBR322 DNA and cauliflower DNA topoisomerase, as assayed by supercoiling of relaxed circular DNA and by digestion with micrococcal nuclease. The optimum ionic strength for supercoiling was 150 mM KCl and the optimum weight ratio of histone to DNA was approximately 1.0. Four histones, H2A, H2B, H3 and H4, were necessary for the optimum assembling conditions, and the nucleosomes assembled protected DNA fragments of approximately 150 bp in length. It was found that cauliflower DNA topoisomerase acts not only as a DNA-relaxing enzyme but also as a chaperon factor for nucleosome assembly.
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Bonner J, Chalkley GR, Dahmus M, Fambrough D, Fujimura F, Huang RC, Huberman J, Jensen R, Marushige K, Ohlenbusch H, Olivera B, Widholm J: Isolation and characterization of chromosomal nucleo-proteins. Meth Enzymol 12B: 3–65 (1968).
Earnshaw WC, Honda BM, Laskey RA, Thomas JO: Assembly of nucleosomes: the reaction involving Xenopus laevis. Cell 21: 373–383 (1980).
Ellison MJ, Pulleyblank DE: The assembly of an H2A2, H2B2, H3, H4 hexamer onto DNA under conditions of physiological ionic strength. J Biol Chem 258: 13307–13313 (1983).
Felsenfeld G: Chromatin. Nature 271: 115–122 (1978).
Fukasawa H, Hamada Y: Properties of soluble chromatins isolated from apical and axial parts of cauliflower inflorescence. Exp Cell Res 88: 352–358 (1974).
Fukata H, Fukasawa H: Isolation and partial characterization of two distinct DNA topoisomerases from cauli-flower inflorescence. J Biochem 91: 1337–1342 (1982).
Fukata H, Fukasawa H: Enzyme activity and processivity of DNA relaxation of salt concentrations of cauliflower DNA topoisomerase II. Jpn J Genet 59: 303–309 (1984).
Fukata H, Ohgami K, Fukasawa H: Isolation and characterization of DNA topoisomerase II from cauliflower inflorescences. Plant Mol Biol 6: 137–144 (1986).
Fukata H, Fukasawa H: Stimulation of RNA synthesis in isolated chromatin by an addition of DNA topoisomerases from cauliflower inflorescence. Jpn J Genet 62: 39–43 (1987).
Gellert M: DNA topoisomerases. Ann Rev Biochem 50: 879–910 (1981).
Germond JE, Hirt B, Oudet P, Gross-Bellard M, Chambon P: Folding of the DNA double helix in chromatin-like structures from simian virus 40. Proc Natl Acad Sci USA 72: 1843–1847 (1975).
Germond JE, Rouviere-Yaniv J, Yaniv M, Brutlag D: Nicking-closing enzyme assembles nucleosome-like structures in vitro. Proc Natl Acad Sci USA 76: 3779–3783 (1979).
Goodwin GH, Sanders C, Johns EW: A new group of chromatin-associated proteins with a high content of acidic and basic amino acids. Eur J Biochem 38: 14–19 (1973).
Ishimi Y, Hirosumi J, Sato W, Sugasawa K, Yokota S, Hanaoka F, Yamada M: Purification and initial characterization of a protein which facilitates assembly of nucleosome-like structure from mammalian cells. Eur J Biochem 142: 431–439 (1984).
Itzhaki RF, Gill DM: A micro-biuret method for estimating proteins. Anal Biochem 9: 401–410 (1964).
Kornberg RD: Structure of chromatin. Ann Rev Biochem 46: 931–954 (1977).
Laskey RA, Mills AD, Morris NR: Assembly of SV40 chromatin in a cell-free system from Xenopus eggs. Cell 10: 237–243 (1977).
Laskey RA, Honda BM, Mills AD, Finch JT: Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA. Nature 275: 416–420 (1978).
Laskey RA, Earnshaw WC: Nucleosome assembly. Nature 286: 763–767 (1980).
Nadeau P, Pallotta D, LaFontaine J: Electrophoretic study of plant histones: Comparison with vertebrate histones. Arch Biochem Biophys 161: 171–177 (1974).
Nelson T. Wiegand R, Brutlag D: Ribonucleic acid and other polyanions facilitate chromatin assembly in vitro. Biochemistry 20: 2594–2601 (1981).
Ruiz-Carrillo A, Jorcano JL, Eder G, Lurz R: In vitro core particle and nucleosome assembly at physiological ionic strength. Proc Natl Acad Sci USA 76: 3284–3288 (1979).
Spiker S: Histone variants in plants. Evidence for primary structure variants differing in molecular weight. J Biol Chem 257: 14250–14255 (1982).
Stein A, Whitlock JPJr., Bina M: Acidic polypeptides can assemble both histones and chromatin in vitro at physiological ionic strength. Proc Natl Acad Sci USA 76: 5000–5004 (1979).
Thomas JO, Kornberg RD: An octamer of histones in chromatin and free in solution. Proc Natl Acad Sci USA 72: 2626–2630 (1975).
Wang JC, Liu LF: DNA topoisomerases: Enzymes that catalyze the concerted breaking and rejoining of DNA backbone bonds. In: Taylor JH (ed) Molecular Genetics, Part III, pp. 65–88 Academic Press, New York (1979).
Wang JC: DNA topoisomerases. Ann Rev Biochem 54: 655–697 (1985).
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Fukata, H., Ohgami, K. & Fukasawa, H. Assembly of nucleosome-like structures mediated by cauliflower DNA topoisomerase. Plant Mol Biol 12, 601–607 (1989). https://doi.org/10.1007/BF00044151
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DOI: https://doi.org/10.1007/BF00044151