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. 1978 Feb;5(2):505–521. doi: 10.1093/nar/5.2.505

Reconstitution of chromatin: assembly of the nucleosome.

F X Wilhelm, M L Wilhelm, M Erard, M P Duane
PMCID: PMC341998  PMID: 634796

Abstract

The order of reassociation of the four histones H2a, H2b, H3 and H4 to the DNA during the reconstitution of chromatin was determined. At each step of the reconstitution the DNA and associated histones were separated from the free histones by centrifugation in a glycerol gradient. The unbound and reassociated histones were analysed by gel electrophoresis and the histone-DNA complexes characterized by circular dichroism and electron microscopy. We show that H3 and H4 bind first to the DNA between 1.2 M NaCl and 0.85 M NaCl and impose a nucleosome like structure; in a second step histones H2a and H2b are placed around this kernel to complete the nucleosome.

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Selected References

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

  1. Axel R., Melchior W., Jr, Sollner-Webb B., Felsenfeld G. Specific sites of interaction between histones and DNA in chromatin. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4101–4105. doi: 10.1073/pnas.71.10.4101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bina-Stein M., Simpson R. T. Specific folding and contraction of DNA by histones H3 and H4. Cell. 1977 Jul;11(3):609–618. doi: 10.1016/0092-8674(77)90078-2. [DOI] [PubMed] [Google Scholar]
  3. Boseley P. G., Bradbury E. M., Butler-Browne G. S., Carpenter B. G., Stephens R. M. Physical studies of chromatin. The recombination of histones with DNA. Eur J Biochem. 1976 Feb 2;62(1):21–31. doi: 10.1111/j.1432-1033.1976.tb10093.x. [DOI] [PubMed] [Google Scholar]
  4. Burton D. R., Hyde J. E., Walker I. O. Histones F2a1 and F3 interact reversibly and cooperatively with DNA to form an equimolar complex in chromatin. FEBS Lett. 1975 Jul 15;55(1):77–80. doi: 10.1016/0014-5793(75)80962-8. [DOI] [PubMed] [Google Scholar]
  5. Camerini-Otero R. D., Felsenfeld G. Supercoiling energy and nucleosome formation: the role of the arginine-rich histone kernel. Nucleic Acids Res. 1977;4(5):1159–1181. doi: 10.1093/nar/4.5.1159-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Camerini-Otero R. D., Sollner-Webb B., Felsenfeld G. The organization of histones and DNA in chromatin: evidence for an arginine-rich histone kernel. Cell. 1976 Jul;8(3):333–347. doi: 10.1016/0092-8674(76)90145-8. [DOI] [PubMed] [Google Scholar]
  7. D'Anna J. A., Jr, Isenberg I. Interaction of renatured histones f3 and f2al. Biochem Biophys Res Commun. 1974 Nov 6;61(1):343–347. doi: 10.1016/0006-291x(74)90572-5. [DOI] [PubMed] [Google Scholar]
  8. Dubochet J., Ducommun M., Zollinger M., Kellenberger E. A new preparation method for dark-field electron microscopy of biomacromolecules. J Ultrastruct Res. 1971 Apr;35(1):147–167. doi: 10.1016/s0022-5320(71)80148-x. [DOI] [PubMed] [Google Scholar]
  9. Finch J. T., Lutter L. C., Rhodes D., Brown R. S., Rushton B., Levitt M., Klug A. Structure of nucleosome core particles of chromatin. Nature. 1977 Sep 1;269(5623):29–36. doi: 10.1038/269029a0. [DOI] [PubMed] [Google Scholar]
  10. Gadski R. A., Chae C. B. Mode of reconstitution of chicken erythrocyte and reticulocyte chromatin. Biochemistry. 1976 Aug 24;15(17):3812–3817. doi: 10.1021/bi00662a025. [DOI] [PubMed] [Google Scholar]
  11. Garel A., Kovacs A. M., Champagne M., Daune M. Circular dichroism as a probe of DNA structure inside reconstituted nucleohistones. Nucleic Acids Res. 1976 Oct;3(10):2507–2519. doi: 10.1093/nar/3.10.2507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Germond J. E., Bellard M., Oudet P., Chambon P. Stability of nucleosomes in native and reconstituted chromatins. Nucleic Acids Res. 1976 Nov;3(11):3173–3192. doi: 10.1093/nar/3.11.3173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hewish D. R., Burgoyne L. A. Chromatin sub-structure. The digestion of chromatin DNA at regularly spaced sites by a nuclear deoxyribonuclease. Biochem Biophys Res Commun. 1973 May 15;52(2):504–510. doi: 10.1016/0006-291x(73)90740-7. [DOI] [PubMed] [Google Scholar]
  14. Kornberg R. D., Thomas J. O. Chromatin structure; oligomers of the histones. Science. 1974 May 24;184(4139):865–868. doi: 10.1126/science.184.4139.865. [DOI] [PubMed] [Google Scholar]
  15. Laskey R. A., Mills A. D., Morris N. R. Assembly of SV40 chromatin in a cell-free system from Xenopus eggs. Cell. 1977 Feb;10(2):237–243. doi: 10.1016/0092-8674(77)90217-3. [DOI] [PubMed] [Google Scholar]
  16. Olins A. L., Carlson R. D., Olins D. E. Visualization of chromatin substructure: upsilon bodies. J Cell Biol. 1975 Mar;64(3):528–537. doi: 10.1083/jcb.64.3.528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Roark D. E., Geoghegan T. E., Keller G. H. A two-subunit histone complex from calf thymus. Biochem Biophys Res Commun. 1974 Jul 24;59(2):542–547. doi: 10.1016/s0006-291x(74)80014-8. [DOI] [PubMed] [Google Scholar]
  19. Sollner-Webb B., Camerini-Otero R. D., Felsenfeld G. Chromatin structure as probed by nucleases and proteases: evidence for the central role of histones H3 and H4. Cell. 1976 Sep;9(1):179–193. doi: 10.1016/0092-8674(76)90063-5. [DOI] [PubMed] [Google Scholar]
  20. Stein A., Bina-Stein M., Simpson R. T. Crosslinked histone octamer as a model of the nucleosome core. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2780–2784. doi: 10.1073/pnas.74.7.2780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thomas G. J., Jr, Prescott B., Olins D. E. Secondary structure of histones and DNA in chromatin. Science. 1977 Jul 22;197(4301):385–388. doi: 10.1126/science.560060. [DOI] [PubMed] [Google Scholar]
  22. Thomas J. O., Kornberg R. D. An octamer of histones in chromatin and free in solution. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2626–2630. doi: 10.1073/pnas.72.7.2626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weintraub H., Palter K., Van Lente F. Histones H2a, H2b, H3, and H4 form a tetrameric complex in solutions of high salt. Cell. 1975 Sep;6(1):85–110. doi: 10.1016/0092-8674(75)90077-x. [DOI] [PubMed] [Google Scholar]
  24. Wilhelm F. X., Champagne M. H., Daune M. P. Conformation du DNA dans la nucléoprotéine. Eur J Biochem. 1970 Aug;15(2):321–330. doi: 10.1111/j.1432-1033.1970.tb01010.x. [DOI] [PubMed] [Google Scholar]
  25. Wilhelm X., Champagne M. Dissociation de la nucléoprotéine d'érythrocytes de poulets par les sels. Eur J Biochem. 1969 Aug;10(1):102–109. [PubMed] [Google Scholar]
  26. Woodcock C. L. Reconstitution of chromatin subunits. Science. 1977 Mar 25;195(4284):1350–1352. doi: 10.1126/science.841333. [DOI] [PubMed] [Google Scholar]

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