Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1978 Mar;5(3):739–750. doi: 10.1093/nar/5.3.739

Fractionation of chromatin by differential solubility in dilute salt.

R F Itzhaki, A Hell, G D Birnie
PMCID: PMC342020  PMID: 643617

Abstract

Chromatin prepared from the livers of rats was fractionated on the basis of solubility in dilute NaCl. Neither of the fractions obtained was enriched in newly synthesized DNA. The salt-soluble fraction had a higher protein content (usually up to 50%) relative to the DNA, and contained 72% or more of the rapidly synthesized RNA. This RNA was found to be complexed with the salt-soluble deoxyribonucleoprotein, not merely co-solubilized with it. Also, polylysine-binding studies showed that about 70% or more of the nucleic acid phosphates were accessible as compared to about 40% in the unfractionated chromatin. These properties suggested that the soluble fraction was enriched in activity transcribed chromatin. In contrast molecular hybridization studies showed that the complexity of the DNA and its homology with cDNA transcribed from rat-liver polysomal mRNA were the same as those of DNA from unfractionated chromatin, or from the salt-insoluble fraction. This suggests that the criteria commonly accepted as distinguishing between euchromatin and heterochromatin in vitro are not invariably valid.

Full text

PDF
739

Selected References

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

  1. Arnold E. A., Young K. E. Heterogeneity of chromatin: fractionation of sonicated rat liver chromatin by partial precipitation with Mg2+. Arch Biochem Biophys. 1974 Sep;164(1):73–89. doi: 10.1016/0003-9861(74)90009-5. [DOI] [PubMed] [Google Scholar]
  2. Birnie G. D., MacPhail E., Young B. D., Getz M. J., Paul J. The diversity of the messenger RNA population in growing Friend cells. Cell Differ. 1974 Nov;3(4):221–232. doi: 10.1016/0045-6039(74)90005-0. [DOI] [PubMed] [Google Scholar]
  3. Doenecke D., McCarthy B. J. Protein content of chromatin fractions separated by sucrose gradient centrifugation. Biochemistry. 1975 Apr 8;14(7):1366–1372. doi: 10.1021/bi00678a004. [DOI] [PubMed] [Google Scholar]
  4. Elgin S. C., Weintraub H. Chromosomal proteins and chromatin structure. Annu Rev Biochem. 1975;44:725–774. doi: 10.1146/annurev.bi.44.070175.003453. [DOI] [PubMed] [Google Scholar]
  5. Fox B. W. The application of Triton X 100 colloid scintillation counting in biochemistry. Int J Appl Radiat Isot. 1968 Oct;19(10):717–730. doi: 10.1016/0020-708x(68)90109-9. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Harrison P. R., Birnie G. D., Hell A., Humphries S., Young B. D., Paul J. Kinetic studies of gene frequency. I. Use of a DNA copy of reticulocyte 9 S RNA to estimate globin gene dosage in mouse tissues. J Mol Biol. 1974 Apr 25;84(4):539–554. doi: 10.1016/0022-2836(74)90115-6. [DOI] [PubMed] [Google Scholar]
  8. Hell A., Birnie G. D., Slimming T. K., Paul J. Controlled fragmentation of DNA by DNase I. Anal Biochem. 1972 Aug;48(2):369–377. doi: 10.1016/0003-2697(72)90089-9. [DOI] [PubMed] [Google Scholar]
  9. ITZHAKI R. F., GILL D. M. A MICRO-BIURET METHOD FOR ESTIMATING PROTEINS. Anal Biochem. 1964 Dec;9:401–410. doi: 10.1016/0003-2697(64)90200-3. [DOI] [PubMed] [Google Scholar]
  10. Itzhaki R. F. Accessible DNA in chromatin. Eur J Biochem. 1974 Aug 15;47(1):27–33. doi: 10.1111/j.1432-1033.1974.tb03664.x. [DOI] [PubMed] [Google Scholar]
  11. Itzhaki R. F., Cooper H. K. Similarity of chromatin from different tissues. J Mol Biol. 1973 Mar 25;75(1):119–128. doi: 10.1016/0022-2836(73)90533-0. [DOI] [PubMed] [Google Scholar]
  12. Itzhaki R. F. Structure and properties of rat thymus deoxyribonucleoprotein. Solubility properties and effects of precipitation. Biochem J. 1967 Nov;105(2):741–748. doi: 10.1042/bj1050741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Itzhaki R. F. Studies on the accessibility of deoxyribonucleic acid in deoxyribonucleoprotein to cationic molecules. Biochem J. 1971 May;122(4):583–592. doi: 10.1042/bj1220583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Lewin B. Units of transcription and translation: sequence components of heterogeneous nuclear RNA and messenger RNA. Cell. 1975 Feb;4(2):77–93. doi: 10.1016/0092-8674(75)90113-0. [DOI] [PubMed] [Google Scholar]
  16. Paul J., Malcolm S. A class of chromatin particles associated with nonhistone proteins. Biochemistry. 1976 Aug 10;15(16):3510–3515. doi: 10.1021/bi00661a018. [DOI] [PubMed] [Google Scholar]
  17. Rickwood D., Hell A., Malcolm S., Birnie G. D., MacGillivray A. J., Paul J. Fractionation of unfixed chromatin by buoyant-density centrifugation. Biochim Biophys Acta. 1974 Jul 11;353(3):353–361. doi: 10.1016/0005-2787(74)90029-x. [DOI] [PubMed] [Google Scholar]
  18. Seale R. L. Assembly of DNA and protein during replication in HeLa cells. Nature. 1975 May 15;255(5505):247–249. doi: 10.1038/255247a0. [DOI] [PubMed] [Google Scholar]
  19. Simpson R. T. Structure and function of chromatin. Adv Enzymol Relat Areas Mol Biol. 1973;38:41–108. doi: 10.1002/9780470122839.ch2. [DOI] [PubMed] [Google Scholar]
  20. Unuma T., Arendell J. P., Busch H. High resolution autoradiographic studies of the uptake of 3H-5-uridine into condensed and dispersed chromatin of nuclei and granular and fibrillar components of nucleoli of Novikoff hepatoma ascites cells. Exp Cell Res. 1968 Oct;52(2):429–438. doi: 10.1016/0014-4827(68)90484-9. [DOI] [PubMed] [Google Scholar]
  21. Weintraub H. The assembly of newly replicated DNA into chromatin. Cold Spring Harb Symp Quant Biol. 1974;38:247–256. doi: 10.1101/sqb.1974.038.01.028. [DOI] [PubMed] [Google Scholar]
  22. Wetmur J. G., Davidson N. Kinetics of renaturation of DNA. J Mol Biol. 1968 Feb 14;31(3):349–370. doi: 10.1016/0022-2836(68)90414-2. [DOI] [PubMed] [Google Scholar]
  23. Young B. D., Birnie G. D. Complexity and specificity of polysomal poly(A+) RNA in mouse tissues. Biochemistry. 1976 Jun 29;15(13):2823–2829. doi: 10.1021/bi00658a019. [DOI] [PubMed] [Google Scholar]

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

RESOURCES