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. 1978 Jan;5(1):71–85. doi: 10.1093/nar/5.1.71

Proximity and accessibility studies of histones in nuclei and free nucleosomes.

W M Bonner
PMCID: PMC341962  PMID: 643611

Abstract

Histone proximity in chromatin was studied with the cleavable crosslinking reagent, dithiobissuccinimidyl propionate. Crosslinks between H4 and H2a, H4 and H2b, H4 and H3, H2a and H2b, H2b and H3 were found. H1 is also crosslinked to the nucleosomal histones. In nuclei, unsheared chromatin, and H1 depleted chromatin, the four nucleosomal histones are crosslinked at similar relative rates both in 5 mM salt and 100 mM salt. After micrococcal nuclease treatment to generate nucleosomes, H2a and H2b are crosslinked faster than H4 and H3. C14-NEM titration of thiopropionate residues bound to each histone shows that H2a and H2b are more accessible to this reagent after nuclease treatment but that the increased binding was not sufficient by itself to explain the increase in crosslinking. Bolton Hunter reagent was used to further study the accessibility of the four nucleosomal histones in whole chromatin and nuclease digested chromatin. These studies showed that salt increases the accessibility of all four histones while nuclease treatment decreases H4 accessibility.

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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. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Bonner W. M., Pollard H. B. The presence of F3-F2a1 dimers and F1 oligomers in chromatin. Biochem Biophys Res Commun. 1975 May 5;64(1):282–288. doi: 10.1016/0006-291x(75)90250-8. [DOI] [PubMed] [Google Scholar]
  4. Bonner W. M. Protein migration into nuclei. I. Frog oocyte nuclei in vivo accumulate microinjected histones, allow entry to small proteins, and exclude large proteins. J Cell Biol. 1975 Feb;64(2):421–430. doi: 10.1083/jcb.64.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burgoyne L. A., Hewish D. R., Mobbs J. Mammalian chromatin substructure studies with the calcium-magnesium endonuclease and two-dimensional polyacrylamide-gel electrophoresis. Biochem J. 1974 Oct;143(1):67–72. doi: 10.1042/bj1430067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark R. J., Felsenfeld G. Structure of chromatin. Nat New Biol. 1971 Jan 27;229(4):101–106. doi: 10.1038/newbio229101a0. [DOI] [PubMed] [Google Scholar]
  7. Finch J. T., Noll M., Kornberg R. D. Electron microscopy of defined lengths of chromatin. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3320–3322. doi: 10.1073/pnas.72.9.3320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Griffith J. D. Chromatin structure: deduced from a minichromosome. Science. 1975 Mar 28;187(4182):1202–1203. doi: 10.1126/science.187.4182.1202. [DOI] [PubMed] [Google Scholar]
  9. Hardison R. C., Eichner M. E., Chalkley R. An approach to histone nearest neighbours in extended chromatin. Nucleic Acids Res. 1975 Oct;2(10):1751–1770. doi: 10.1093/nar/2.10.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Hyde J. E., Walker I. O. Covalent cross-linking of histones in chromatin. FEBS Lett. 1975 Feb 1;50(2):150–154. doi: 10.1016/0014-5793(75)80477-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Loening U. E. The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem J. 1967 Jan;102(1):251–257. doi: 10.1042/bj1020251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Malchy B., Kaplan H. Reactive properties of the amino groups of histones in calf thymus chromatin. J Mol Biol. 1974 Feb 5;82(4):537–545. doi: 10.1016/0022-2836(74)90247-2. [DOI] [PubMed] [Google Scholar]
  15. Martinson H. G., McCarthy B. J. Histone-histone associations within chromatin. Cross-linking studies using tetranitromethane. Biochemistry. 1975 Mar 11;14(5):1073–1078. doi: 10.1021/bi00676a030. [DOI] [PubMed] [Google Scholar]
  16. Martinson H. G., Shetlar M. D., McCarthy B. J. Histone-histone interactions within chromatin. Crosslinking studies using ultraviolet light. Biochemistry. 1976 May 4;15(9):2002–2007. doi: 10.1021/bi00654a030. [DOI] [PubMed] [Google Scholar]
  17. Noll M. Internal structure of the chromatin subunit. Nucleic Acids Res. 1974 Nov;1(11):1573–1578. doi: 10.1093/nar/1.11.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Olins D. E., Wright E. B. Glutaraldehyde fixation of isolated eucaryotic nuclei. Evidence for histone-histone proximity. J Cell Biol. 1973 Nov;59(2 Pt 1):304–317. doi: 10.1083/jcb.59.2.304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Sahasrabuddhe C. G., Van Holde K. E. The effect of trypsin on nuclease-resistant chromatin fragments. J Biol Chem. 1974 Jan 10;249(1):152–156. [PubMed] [Google Scholar]
  22. Simpson R. T. Histones H3 and H4 interact with the ends of nucleosome DNA. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4400–4404. doi: 10.1073/pnas.73.12.4400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Thomas J. O., Kornberg R. D. Cleavable cross-links in the analysis of histone-histone associations. FEBS Lett. 1975 Oct 15;58(1):353–358. doi: 10.1016/0014-5793(75)80296-1. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Van Lente F., Jackson J. F., Weintraub H. Identification of specific crosslinked histones after treatment of chromatin with formaldehyde. Cell. 1975 May;5(1):45–50. doi: 10.1016/0092-8674(75)90090-2. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. van Bruggen E. F., Arnberg A. C., van Holde K. E., Sahasrabuddhe C. G., Shaw B. R. Electron microscopy of chromatin subunit particles. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1365–1370. doi: 10.1016/0006-291x(74)90348-9. [DOI] [PubMed] [Google Scholar]

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