Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1976 Aug;3(8):2041–2054. doi: 10.1093/nar/3.8.2041

The absence of histone H1 from the chromatin fraction obtained by sonication of calf thymus nuclei under "quasiphysiological" ionic conditions.

A I Lishanskaya, M I Mosevitsky
PMCID: PMC343060  PMID: 967688

Abstract

The minor chromatin fraction was isolated from the sonicated calf thymus nuclei on the basis of its differential solubility in the "quasiphysiological" salt medium (0.1 M KCl-0.05 M NaCl-l mM MgCl2-1 mM CaCl2). Histone Hl is almost completely absent from this fraction. DNA isolated from this fraction occurs in three discrete low mol. wt. fragments. The fraction of chromatin which lacks histone Hl can also be obtained by two other methods. On of them consists in salt precipitation of the chromatin gel and its subsequent sonication. The second method includes precipitation of the sonicated chromatin gel by salts. In the first case the properties of the chromatin fraction which remains in the supernatant after centrifugation closely resemble those of the original salt-soluble nuclear fraction. The second method yields supernatant fraction also lacking histone Hl but containing heterogeneous DNA. Comparisons were also made of the sonically-solubilized nuclear fractions obtained in the complete salt medium and its mono and divalent cationic constituents.

Full text

PDF
2041

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berkowitz E. M., Doty P. Chemical and physical properties of fractionated chromatin. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3328–3332. doi: 10.1073/pnas.72.9.3328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonner J., Gottesfeld J., Garrard W., Billing R., Uphouse L. Isolation of template active and inactive regions of chromatin. Methods Enzymol. 1975;40:97–102. doi: 10.1016/s0076-6879(75)40010-6. [DOI] [PubMed] [Google Scholar]
  3. Chalkley R., Jensen R. H. A study of the structure of isolated chromatin. Biochemistry. 1968 Dec;7(12):4380–4388. doi: 10.1021/bi00852a034. [DOI] [PubMed] [Google Scholar]
  4. FRENSTER J. H., ALLFREY V. G., MIRSKY A. E. REPRESSED AND ACTIVE CHROMATIN ISOLATED FROM INTERPHASE LYMPHOCYTES. Proc Natl Acad Sci U S A. 1963 Dec;50:1026–1032. doi: 10.1073/pnas.50.6.1026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gottesfeld J. M., Garrard W. T., Bagi G., Wilson R. F., Bonner J. Partial purification of the template-active fraction of chromatin: a preliminary report. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2193–2197. doi: 10.1073/pnas.71.6.2193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gottesfeld J. M., Murphy R. F., Bonner J. Structure of transcriptionally active chromatin. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4404–4408. doi: 10.1073/pnas.72.11.4404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lishanskaya A. I., Mosevitsky M. I. Study of calf thymus deoxyribonucleoproteins by means of gel electrophoresis. Effect of ionic composition on the mode of chromatin fragmentation. Biochem Biophys Res Commun. 1975 Feb 17;62(4):822–829. doi: 10.1016/0006-291x(75)90396-4. [DOI] [PubMed] [Google Scholar]
  8. Littau V. C., Burdick C. J., Allfrey V. G., Mirsky S. A. The role of histones in the maintenance of chromatin structure. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1204–1212. doi: 10.1073/pnas.54.4.1204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Marushige K., Bonner J. Fractionation of liver chromatin. Proc Natl Acad Sci U S A. 1971 Dec;68(12):2941–2944. doi: 10.1073/pnas.68.12.2941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McConaughy B. L., McCarthy B. J. Fractionation of chromatin by thermal chromatography. Biochemistry. 1972 Mar 14;11(6):998–1003. doi: 10.1021/bi00756a008. [DOI] [PubMed] [Google Scholar]
  11. Oudet P., Gross-Bellard M., Chambon P. Electron microscopic and biochemical evidence that chromatin structure is a repeating unit. Cell. 1975 Apr;4(4):281–300. doi: 10.1016/0092-8674(75)90149-x. [DOI] [PubMed] [Google Scholar]
  12. Panyim S., Chalkley R. High resolution acrylamide gel electrophoresis of histones. Arch Biochem Biophys. 1969 Mar;130(1):337–346. doi: 10.1016/0003-9861(69)90042-3. [DOI] [PubMed] [Google Scholar]
  13. Reeck G. R., Simpson R. T., Sober H. A. Resolution of a spectrum of nucleoprotein species in sonicated chromatin. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2317–2321. doi: 10.1073/pnas.69.8.2317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Reeck G. R., Simpson R. T., Sober H. A. The distribution of histones and nonhistone proteins in the ECTHAM-cellulose fractions of chromatin from several tissues. Eur J Biochem. 1974 Nov 15;49(2):407–414. doi: 10.1111/j.1432-1033.1974.tb03846.x. [DOI] [PubMed] [Google Scholar]
  15. Samarina O. P., Lukanidin E. M., Molnar J., Georgiev G. P. Structural organization of nuclear complexes containing DNA-like RNA. J Mol Biol. 1968 Apr 14;33(1):251–263. doi: 10.1016/0022-2836(68)90292-1. [DOI] [PubMed] [Google Scholar]
  16. Van Holde K. E., Sahasrabuddhe C. G., Shaw B. R. A model for particulate structure in chromatin. Nucleic Acids Res. 1974 Nov;1(11):1579–1586. doi: 10.1093/nar/1.11.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Varshavsky A. J., Bakayev V. V. Studies on chromatin. III. v-Bodies and free DNA in chromatin lacking histone H1. Mol Biol Rep. 1975 Oct;2(3):209–217. doi: 10.1007/BF00356990. [DOI] [PubMed] [Google Scholar]
  18. Warnecke P., Kruse K., Harbers E. Studies on deoxyribonucleoproteins. 8. Isolation and characterization of nonhistone proteins from euchromatic and heterochromatic deoxyribonucleoprotein of rat liver. Biochim Biophys Acta. 1973 Dec 7;331(2):295–304. [PubMed] [Google Scholar]
  19. Yasmineh W. G., Yunis J. J. Satellite DNA in mouse autosomal heterochromatin. Biochem Biophys Res Commun. 1969 Jun 27;35(6):779–782. doi: 10.1016/0006-291x(69)90690-1. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES