Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Sep;72(9):3328–3332. doi: 10.1073/pnas.72.9.3328

Chemical and physical properties of fractionated chromatin.

E M Berkowitz, P Doty
PMCID: PMC432985  PMID: 1059117

Abstract

Sonicated chicken reticulocyte chromatin was fractionated into transcriptionally active and transcriptionally repressed components. The active fraction is 8% of the whole chromatin but contains 70% of the newly synthesized chromosomal RNA. This RNA has five times as many hemoglobin RNA sequences as does the RNA in the repressed fraction. The amount of the active fraction in the chromatins of several tissues correlates with their synthetic activity. The molecular weight of the DNA of the repressed fraction is approximately twice that of the active fraction. Moreover, the configuration of repressed chromatin is much more compact, consistent with a much larger sedimentation constant. The transcriptionally active fraction displays a 6 degrees lower melting profile and is highly susceptible to DNase I relative to the repressed fraction. The active fraction contains twice as much non-histone protein and 15% less histone than the repressed fraction and is lacking the lysine-rich and much of the arginine-rich histones.

Full text

PDF
3328

Selected References

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

  1. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonner J., Garrard W. T., Gottesfeld J., Holmes D. S. Functional organization of the mammalian genome. Cold Spring Harb Symp Quant Biol. 1974;38:303–310. doi: 10.1101/sqb.1974.038.01.034. [DOI] [PubMed] [Google Scholar]
  3. Burgess R. R. A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase. J Biol Chem. 1969 Nov 25;244(22):6160–6167. [PubMed] [Google Scholar]
  4. Cedar H., Felsenfeld G. Transcription of chromatin in vitro. J Mol Biol. 1973 Jun 25;77(2):237–254. doi: 10.1016/0022-2836(73)90334-3. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Comings D. E., Mattoccia E. DNA of mammalian and avian heterochromatin. Exp Cell Res. 1972 Mar;71(1):113–131. doi: 10.1016/0014-4827(72)90270-4. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Duerksen J. D., McCarthy B. J. Distribution of deoxyribonucleic acid sequences in fractionated chromatin. Biochemistry. 1971 Apr 13;10(8):1471–1478. doi: 10.1021/bi00784a031. [DOI] [PubMed] [Google Scholar]
  9. Elgin S. C., Bonner J. Limited heterogeneity of the major nonhistone chromosomal proteins. Biochemistry. 1970 Oct 27;9(22):4440–4447. doi: 10.1021/bi00824a027. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Frenster J. H. Nuclear polyanions as de-repressors of synthesis of ribonucleic acid. Nature. 1965 May 15;206(985):680–683. doi: 10.1038/206680a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Howk R. S., Anisowicz A., Silverman A. Y., Parks W. P., Scoinick E. M. Distribution of murine type B and type C viral nucleic acid sequences in template active and template inactive chromatin. Cell. 1975 Apr;4(4):321–327. doi: 10.1016/0092-8674(75)90152-x. [DOI] [PubMed] [Google Scholar]
  14. Kornberg R. D. Chromatin structure: a repeating unit of histones and DNA. Science. 1974 May 24;184(4139):868–871. doi: 10.1126/science.184.4139.868. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lin R. I., Schjeide O. A. Micro estimation of RNA by the cupric ion catalyzed orcinol reaction. Anal Biochem. 1969 Mar;27(3):473–483. doi: 10.1016/0003-2697(69)90061-x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. McCarthy B. J., Nishiura J. T., Doenecke D., Nasser D. S., Johnson C. B. Transcription and chromatin structure. Cold Spring Harb Symp Quant Biol. 1974;38:763–771. doi: 10.1101/sqb.1974.038.01.081. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Murphy E. C., Jr, Hall S. H., Shepherd J. H., Weiser R. S. Fractionation of mouse myeloma chromatin. Biochemistry. 1973 Sep 25;12(20):3843–3853. doi: 10.1021/bi00744a008. [DOI] [PubMed] [Google Scholar]
  21. Noll M. Subunit structure of chromatin. Nature. 1974 Sep 20;251(5472):249–251. doi: 10.1038/251249a0. [DOI] [PubMed] [Google Scholar]
  22. Olins A. L., Olins D. E. Spheroid chromatin units (v bodies). Science. 1974 Jan 25;183(4122):330–332. doi: 10.1126/science.183.4122.330. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Preisler H. S., Housman D., Scher W., Friend C. Effects of 5-bromo-2' -deoxyuridine on production of globin messenger RNA in dimethyl sulfoxide-stimulated Friend leukemia cells. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2956–2959. doi: 10.1073/pnas.70.10.2956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Simpson R. T. Distribution of RNA polymerase binding sites in fractionated chromatin. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2740–2743. doi: 10.1073/pnas.71.7.2740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Simpson R. T., Reeck G. R. A comparison of the proteins of condensed and extended chromatin fractions of rabbit liver and calf thymus. Biochemistry. 1973 Sep 25;12(20):3853–3858. doi: 10.1021/bi00744a009. [DOI] [PubMed] [Google Scholar]
  29. Varshavsky A. J., Ilyin Y. V., Georgiev G. P. Very long stretches of free DNA in chromatin. Nature. 1974 Aug 16;250(467):602–606. doi: 10.1038/250602a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES