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. 1986 Mar;5(3):519–528. doi: 10.1002/j.1460-2075.1986.tb04241.x

Cryo-electron microscopy of vitrified SV40 minichromosomes: the liquid drop model.

J Dubochet, M Adrian, P Schultz, P Oudet
PMCID: PMC1166793  PMID: 3011410

Abstract

The structure of SV40 minichromosomes has been studied by cryo-electron microscopy of vitrified thin layers of solution. In high-salt buffer (130 mM NaCl), freshly prepared minichromosomes are condensed into globules 30 nm or more in diameter. On the micrograph, they appear to be formed by the close packing of 10 nm granules which give rise to a 10 nm reflection in the optical diffractogram. The globules can adopt many different conformations. At high concentration, they fuse into a homogeneous 'sea' of closely packed 10 nm granules. In low-salt buffer (less than 10 mM NaCl), the globules open, first into 10 nm filaments, and then into nucleosome-strings. The 'liquid drop' model is proposed to explain the condensed structure of the minichromosome in high-salt buffer: nucleosomes stack specifically on top of one another, thus forming the 10 nm filaments. 10 nm filaments in turn, tend to aggregate laterally. Optimizing both these interactions results in the condensation of 10 nm filaments or portions thereof into a structure similar to that of a liquid. Some implications of this model for the structure of cellular chromatin are discussed.

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Selected References

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  1. Adrian M., Dubochet J., Lepault J., McDowall A. W. Cryo-electron microscopy of viruses. Nature. 1984 Mar 1;308(5954):32–36. doi: 10.1038/308032a0. [DOI] [PubMed] [Google Scholar]
  2. Bellard M., Oudet P., Germond J. E., Chambon P. Subunit structure of simian-virus-40 minichromosome. Eur J Biochem. 1976 Nov 15;70(2):543–553. doi: 10.1111/j.1432-1033.1976.tb11046.x. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Dubochet J., Noll M. Nucleosome arcs and helices. Science. 1978 Oct 20;202(4365):280–286. doi: 10.1126/science.694532. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Jakobovits E. B., Bratosin S., Aloni Y. A nucleosome-free region in SV40 minichromosomes. Nature. 1980 May 22;285(5762):263–265. doi: 10.1038/285263a0. [DOI] [PubMed] [Google Scholar]
  8. Jongstra J., Reudelhuber T. L., Oudet P., Benoist C., Chae C. B., Jeltsch J. M., Mathis D. J., Chambon P. Induction of altered chromatin structures by simian virus 40 enhancer and promoter elements. Nature. 1984 Feb 23;307(5953):708–714. doi: 10.1038/307708a0. [DOI] [PubMed] [Google Scholar]
  9. Keller W., Müller U., Eicken I., Wendel I., Zentgraf H. Biochemical and ultrastructural analysis of SV40 chromatin. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):227–244. doi: 10.1101/sqb.1978.042.01.025. [DOI] [PubMed] [Google Scholar]
  10. Lepault J., Booy F. P., Dubochet J. Electron microscopy of frozen biological suspensions. J Microsc. 1983 Jan;129(Pt 1):89–102. doi: 10.1111/j.1365-2818.1983.tb04163.x. [DOI] [PubMed] [Google Scholar]
  11. Lepault J., Pitt T. Projected structure of unstained, frozen-hydrated T-layer of Bacillus brevis. EMBO J. 1984 Jan;3(1):101–105. doi: 10.1002/j.1460-2075.1984.tb01768.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Livolant F. Cholesteric organization of DNA in vivo and in vitro. Eur J Cell Biol. 1984 Mar;33(2):300–311. [PubMed] [Google Scholar]
  13. McDowall A. W., Hofmann W., Lepault J., Adrian M., Dubochet J. Cryo-electron microscopy of vitrified insect flight muscle. J Mol Biol. 1984 Sep 5;178(1):105–111. doi: 10.1016/0022-2836(84)90233-x. [DOI] [PubMed] [Google Scholar]
  14. Mirkovitch J., Mirault M. E., Laemmli U. K. Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold. Cell. 1984 Nov;39(1):223–232. doi: 10.1016/0092-8674(84)90208-3. [DOI] [PubMed] [Google Scholar]
  15. Moyne G., Harper F., Saragosti S., Yaniv M. Absence of nucleosomes in a histone-containing nucleoprotein complex obtained by dissociation of purified SV40 virions. Cell. 1982 Aug;30(1):123–130. doi: 10.1016/0092-8674(82)90018-6. [DOI] [PubMed] [Google Scholar]
  16. Müller U., Zentgraf H., Eicken I., Keller W. Higher order structure of simian virus 40 chromatin. Science. 1978 Aug 4;201(4354):406–415. doi: 10.1126/science.208155. [DOI] [PubMed] [Google Scholar]
  17. Noll M., Thomas J. O., Kornberg R. D. Preparation of native chromatin and damage caused by shearing. Science. 1975 Mar 28;187(4182):1203–1206. doi: 10.1126/science.187.4182.1203. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Richmond T. J., Finch J. T., Rushton B., Rhodes D., Klug A. Structure of the nucleosome core particle at 7 A resolution. Nature. 1984 Oct 11;311(5986):532–537. doi: 10.1038/311532a0. [DOI] [PubMed] [Google Scholar]
  20. Saragosti S., Moyne G., Yaniv M. Absence of nucleosomes in a fraction of SV40 chromatin between the origin of replication and the region coding for the late leader RNA. Cell. 1980 May;20(1):65–73. doi: 10.1016/0092-8674(80)90235-4. [DOI] [PubMed] [Google Scholar]
  21. Shelton E. R., Wassarman P. M., DePamphilis M. L. Structure, spacing, and phasing of nucleosomes on isolated forms of mature simian virus 40 chromosomes. J Biol Chem. 1980 Jan 25;255(2):771–782. [PubMed] [Google Scholar]
  22. Thoma F., Koller T., Klug A. Involvement of histone H1 in the organization of the nucleosome and of the salt-dependent superstructures of chromatin. J Cell Biol. 1979 Nov;83(2 Pt 1):403–427. doi: 10.1083/jcb.83.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Varshavsky A. J., Bakayev V. V., Nedospasov S. A., Georgiev G. P. On the structure of eukaryotic, prokaryotic, and viral chromatin. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):457–473. doi: 10.1101/sqb.1978.042.01.049. [DOI] [PubMed] [Google Scholar]
  24. Varshavsky A. J., Nedospasov S. A., Schmatchenko V. V., Bakayev V. V., Chumackov P. M., Georgiev G. P. Compact form of SV40 viral minichromosome is resistant to nuclease: possible implications for chromatin structure. Nucleic Acids Res. 1977 Oct;4(10):3303–3325. doi: 10.1093/nar/4.10.3303. [DOI] [PMC free article] [PubMed] [Google Scholar]

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