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. 1986 Jun;5(6):1395–1402. doi: 10.1002/j.1460-2075.1986.tb04373.x

Cryo-electron microscopy of vitrified chromosomes in situ.

A W McDowall, J M Smith, J Dubochet
PMCID: PMC1166954  PMID: 3755397

Abstract

Chromosomes of metaphase-arrested Chinese hamster ovary (CHO) and HeLa cells were examined in situ, unfixed and unstained, by cryo-electron microscopy. In hydrated, vitrified cryo-sections, chromosomes exhibit a characteristic homogeneous, grainy texture, which, on optical diffraction, gives rise to a broad reflection corresponding to 11 nm. No superstructure or periodic order is discernible. These observations suggest that the chromosome is formed by the compact association of 11 nm filaments, or portions thereof, interacting in a manner akin to the molecules of a liquid. Some implications of the liquid model of chromosome structure are discussed.

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Selected References

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

  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. Butler P. J. The folding of chromatin. CRC Crit Rev Biochem. 1983;15(1):57–91. doi: 10.3109/10409238309102801. [DOI] [PubMed] [Google Scholar]
  3. Compton J. L., Hancock R., Oudet P., Chambon P. Biochemical and electron-microscopic evidence that the subunit structure of Chinese-hamster-ovary interphase chromatin is conserved in mitotic chromosomes. Eur J Biochem. 1976 Nov 15;70(2):555–568. doi: 10.1111/j.1432-1033.1976.tb11047.x. [DOI] [PubMed] [Google Scholar]
  4. Davies H. G., Small J. V. Structural units in chromatin and their orientation on membranes. Nature. 1968 Mar 23;217(5134):1122–1125. doi: 10.1038/2171122a0. [DOI] [PubMed] [Google Scholar]
  5. Dubochet J., Adrian M., Schultz P., Oudet P. Cryo-electron microscopy of vitrified SV40 minichromosomes: the liquid drop model. EMBO J. 1986 Mar;5(3):519–528. doi: 10.1002/j.1460-2075.1986.tb04241.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dubochet J., McDowall A. W., Menge B., Schmid E. N., Lickfeld K. G. Electron microscopy of frozen-hydrated bacteria. J Bacteriol. 1983 Jul;155(1):381–390. doi: 10.1128/jb.155.1.381-390.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Finch J. T., Klug A. Solenoidal model for superstructure in chromatin. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1897–1901. doi: 10.1073/pnas.73.6.1897. [DOI] [PMC free article] [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. Jorcano J. L., Meyer G., Day L. A., Renz M. Aggregation of small oligonucleosomal chains into 300-A globular particles. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6443–6447. doi: 10.1073/pnas.77.11.6443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Langmore J. P., Paulson J. R. Low angle x-ray diffraction studies of chromatin structure in vivo and in isolated nuclei and metaphase chromosomes. J Cell Biol. 1983 Apr;96(4):1120–1131. doi: 10.1083/jcb.96.4.1120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Langmore J. P., Schutt C. The higher order structure of chicken erythrocyte chromosomes in vivo. Nature. 1980 Dec 11;288(5791):620–622. doi: 10.1038/288620a0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Lewis C. D., Laemmli U. K. Higher order metaphase chromosome structure: evidence for metalloprotein interactions. Cell. 1982 May;29(1):171–181. doi: 10.1016/0092-8674(82)90101-5. [DOI] [PubMed] [Google Scholar]
  15. McDowall A. W., Chang J. J., Freeman R., Lepault J., Walter C. A., Dubochet J. Electron microscopy of frozen hydrated sections of vitreous ice and vitrified biological samples. J Microsc. 1983 Jul;131(Pt 1):1–9. doi: 10.1111/j.1365-2818.1983.tb04225.x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Paulson J. R., Laemmli U. K. The structure of histone-depleted metaphase chromosomes. Cell. 1977 Nov;12(3):817–828. doi: 10.1016/0092-8674(77)90280-x. [DOI] [PubMed] [Google Scholar]
  18. Rattner J. B., Hamkalo B. A. Higher order structure in metaphase chromosomes. I. The 250 A fiber. Chromosoma. 1978 Dec 6;69(3):363–372. doi: 10.1007/BF00332139. [DOI] [PubMed] [Google Scholar]
  19. Stillinger F. H. Water revisited. Science. 1980 Jul 25;209(4455):451–457. doi: 10.1126/science.209.4455.451. [DOI] [PubMed] [Google Scholar]
  20. Widom J., Klug A. Structure of the 300A chromatin filament: X-ray diffraction from oriented samples. Cell. 1985 Nov;43(1):207–213. doi: 10.1016/0092-8674(85)90025-x. [DOI] [PubMed] [Google Scholar]
  21. Woodcock C. L., Frado L. L., Rattner J. B. The higher-order structure of chromatin: evidence for a helical ribbon arrangement. J Cell Biol. 1984 Jul;99(1 Pt 1):42–52. doi: 10.1083/jcb.99.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zentgraf H., Franke W. W. Differences of supranucleosomal organization in different kinds of chromatin: cell type-specific globular subunits containing different numbers of nucleosomes. J Cell Biol. 1984 Jul;99(1 Pt 1):272–286. doi: 10.1083/jcb.99.1.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zieve G. W., Turnbull D., Mullins J. M., McIntosh J. R. Production of large numbers of mitotic mammalian cells by use of the reversible microtubule inhibitor nocodazole. Nocodazole accumulated mitotic cells. Exp Cell Res. 1980 Apr;126(2):397–405. doi: 10.1016/0014-4827(80)90279-7. [DOI] [PubMed] [Google Scholar]

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