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Nucleic Acids Res. Mar 11, 1982; 10(5): 1481–1494.
PMCID: PMC320544

Analysis of highly purified satellite DNA containing chromatin from the mouse.


A purification scheme for satellite DNA containing chromatin from mouse liver has been developed. It is based on the highly condensed state of the satellite chromatin and also takes advantage of its resistance to digestion by certain restriction nucleases. Nuclei are first treated with micrococcal nuclease and the satellite chromatin enriched 3-5 fold by extraction of the digested nuclei under appropriate conditions. Further purification is achieved by digestion of the chromatin with a restriction nuclease that leaves satellite DNA largely intact but degrades non-satellite DNA extensively. In subsequent sucrose gradient centrifugation the rapidly sedimenting chromatin contains more than 70% satellite DNA. This material has the same histone composition as bulk chromatin. No significant differences were detected in an analysis of minor histone variants. Nonhistone proteins are present only in very low amounts in the satellite chromatin fraction, notably the HMG proteins are strongly depleted.

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Selected References

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  • Brutlag DL. Molecular arrangement and evolution of heterochromatic DNA. Annu Rev Genet. 1980;14:121–144. [PubMed]
  • Baker WK. Position-effect variegation. Adv Genet. 1968;14:133–169. [PubMed]
  • Horvath P, Hörz W. The compaction of mouse heterochromatin as studied by nuclease digestion. FEBS Lett. 1981 Nov 2;134(1):25–28. [PubMed]
  • Hörz W, Zachau HG. Deoxyribonuclease II as a probe for chromatin structure. I. Location of cleavage sites. J Mol Biol. 1980 Dec 15;144(3):305–327. [PubMed]
  • Takatsuka Y, Kohno M, Higashi K, Hirano H, Sakamoto Y. Redistribution of chromatin containing ribosomal cistrons during liver regeneration. Exp Cell Res. 1976 Nov;103(1):191–199. [PubMed]
  • Igo-Kemenes T, Greil W, Zachau HG. Prepartation of soluble chromatin and specific chromatin fractions with restriction nucleases. Nucleic Acids Res. 1977 Oct;4(10):3387–3400. [PMC free article] [PubMed]
  • Kiss A, Sain B, Csordás-Tòth E, Venetianer P. A new sequence-specific endonuclease (Bsp) from Bacillus sphaericus. Gene. 1977 Jul;1(5-6):323–329. [PubMed]
  • Greene PJ, Heyneker HL, Bolivar F, Rodriguez RL, Betlach MC, Covarrubias AA, Backman K, Russel DJ, Tait R, Boyer HW. A general method for the purification of restriction enzymes. Nucleic Acids Res. 1978 Jul;5(7):2373–2380. [PMC free article] [PubMed]
  • Manuelidis L. A simplified method for preparation of mouse satellite DNA. Anal Biochem. 1977 Apr;78(2):561–568. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Goodwin GH, Nicolas RH, Johns EW. An improved large scale fractionation of high mobility group non-histone chromatin proteins. Biochim Biophys Acta. 1975 Oct 20;405(2):280–291. [PubMed]
  • Johns EW. The isolation and purification of histones. Methods Cell Biol. 1977;16:183–203. [PubMed]
  • Billings PC, Orf JW, Palmer DK, Talmage DA, Pan CG, Blumenfeld M. Anomalous electrophoretic mobility of Drosophila phosphorylated H1 histone: is it related to the compaction of satellite DNA into heterochromatin? Nucleic Acids Res. 1979;6(6):2151–2164. [PMC free article] [PubMed]
  • Zweidler A. Resolution of histones by polyacrylamide gel electrophoresis in presence of nonionic detergents. Methods Cell Biol. 1978;17:223–233. [PubMed]
  • Merril CR, Dunau ML, Goldman D. A rapid sensitive silver stain for polypeptides in polyacrylamide gels. Anal Biochem. 1981 Jan 1;110(1):201–207. [PubMed]
  • Hörz W, Zachau HG. Characterization of distinct segments in mouse satellite DNA by restriction nucleases. Eur J Biochem. 1977 Mar 1;73(2):383–392. [PubMed]
  • von Gabain A, Hayward GS, Bujard H. Physical mapping of the HindIII, EcoRI, Sal and Sma restriction endonuclease cleavage fragments from bacteriophage T5 DNA. Mol Gen Genet. 1976 Feb 2;143(3):279–290. [PubMed]
  • McCarty KS, Jr, Vollmer RT, McCarty KS. Improved computer program data for the resolution and fractionation of macromolecules by isokinetic sucrose density gradient sedimentation. Anal Biochem. 1974 Sep;61(1):165–183. [PubMed]
  • Seale RL. Rapid turnover of the histone-ubiquitin conjugate, protein A24. Nucleic Acids Res. 1981 Jul 10;9(13):3151–3158. [PMC free article] [PubMed]
  • Lipchitz L, Axel R. Restriction endonuclease cleavage of satellite DNA in intact bovine nuclei. Cell. 1976 Oct;9(2):355–364. [PubMed]
  • Omori A, Igo-Kemenes T, Zachau HG. Different repeat lengths in rat satellite I DNA containing chromatin and bulk chromatin. Nucleic Acids Res. 1980 Nov 25;8(22):5363–5375. [PMC free article] [PubMed]
  • Musich PR, Brown FL, Maio JJ. Subunit structure of chromatin and the organization of eukaryotic highly repetitive DNA: nucleosomal proteins associated with a highly repetitive mammalian DNA. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3297–3301. [PMC free article] [PubMed]
  • Bernstine EG. Satellite DNA content of chromatin fractions isolated from Eco R1-digested mouse liver nuclei. Exp Cell Res. 1978 Apr;113(1):205–208. [PubMed]
  • Mazrimas JA, Balhorn R, Hatch FT. Separation of satellite DNA chromatin and main band DNA chromatin from mouse brain. Nucleic Acids Res. 1979 Oct 25;7(4):935–946. [PMC free article] [PubMed]
  • Hyde JE, Igo-Kemenes T, Zachau HG. The non-histone proteins of the rat liver nucleus and their distribution amongst chromatin fractions as produced by nuclease digestion. Nucleic Acids Res. 1979 Sep 11;7(1):31–48. [PMC free article] [PubMed]
  • Mathew CG, Goodwin GH, Igo-Kemenes T, Johns EW. The protein composition of rat satellite chromatin. FEBS Lett. 1981 Mar 9;125(1):25–29. [PubMed]
  • Hsieh T, Brutlag DL. A protein that preferentially binds Drosophila satellite DNA. Proc Natl Acad Sci U S A. 1979 Feb;76(2):726–730. [PMC free article] [PubMed]
  • Zachau HG, Igo-Kemenes T. Face to phase with nucleosomes. Cell. 1981 Jun;24(3):597–598. [PubMed]
  • Mathew CG, Goodwin GH, Johns EW. Quantitative analysis of non-histone chromosomal proteins HMG 14 and HMG 17 by polyacrylamide gel electrophoresis. J Chromatogr. 1980 Sep 26;198(1):80–83. [PubMed]
  • Levinger L, Barsoum J, Varshavsky A. Two-dimensional hybridization mapping of nucleosomes. comparison of DNA and protein patterns. J Mol Biol. 1981 Mar 5;146(3):287–304. [PubMed]

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