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. 1994 Apr 1;125(1):1–10. doi: 10.1083/jcb.125.1.1

The three-dimensional architecture of chromatin in situ: electron tomography reveals fibers composed of a continuously variable zig-zag nucleosomal ribbon

PMCID: PMC2120010  PMID: 8138564

Abstract

The three dimensional (3D) structure of chromatin fibers in sections of nuclei has been determined using electron tomography. Low temperature embedding and nucleic acid-specific staining allowed individual nucleosomes to be clearly seen, and the tomographic data collection parameters provided a reconstruction resolution of 2.5 nm. Chromatin fibers have complex 3D trajectories, with smoothly bending regions interspersed with abrupt changes in direction, and U turns. Nucleosomes are located predominantly at the fiber periphery, and linker DNA tends to project toward the fiber interior. Within the fibers, a unifying structural motif is a two nucleosome-wide ribbon that is variably bent and twisted, and in which there is little face-to-face contact between nucleosomes. It is suggested that this asymmetric 3D zig-zag of nucleosomes and linker DNA represents a basic principle of chromatin folding that is determined by the properties of the nucleosome-linker unit. This concept of chromatin fiber architecture is contrasted with helical models in which specific nucleosome-nucleosome contacts play a major role in generating a symmetrical higher order structure. The transcriptional control implications of a more open and irregular chromatin structure are discussed.

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Selected References

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  1. Armbruster B. L., Carlemalm E., Chiovetti R., Garavito R. M., Hobot J. A., Kellenberger E., Villiger W. Specimen preparation for electron microscopy using low temperature embedding resins. J Microsc. 1982 Apr;126(Pt 1):77–85. doi: 10.1111/j.1365-2818.1982.tb00358.x. [DOI] [PubMed] [Google Scholar]
  2. Athey B. D., Smith M. F., Rankert D. A., Williams S. P., Langmore J. P. The diameters of frozen-hydrated chromatin fibers increase with DNA linker length: evidence in support of variable diameter models for chromatin. J Cell Biol. 1990 Sep;111(3):795–806. doi: 10.1083/jcb.111.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baumeister W., Barth M., Hegerl R., Guckenberger R., Hahn M., Saxton W. O. Three-dimensional structure of the regular surface layer (HPI layer) of Deinococcus radiodurans. J Mol Biol. 1986 Jan 20;187(2):241–250. doi: 10.1016/0022-2836(86)90231-7. [DOI] [PubMed] [Google Scholar]
  4. Belmont A. S., Braunfeld M. B., Sedat J. W., Agard D. A. Large-scale chromatin structural domains within mitotic and interphase chromosomes in vivo and in vitro. Chromosoma. 1989 Aug;98(2):129–143. doi: 10.1007/BF00291049. [DOI] [PubMed] [Google Scholar]
  5. Belmont A. S., Sedat J. W., Agard D. A. A three-dimensional approach to mitotic chromosome structure: evidence for a complex hierarchical organization. J Cell Biol. 1987 Jul;105(1):77–92. doi: 10.1083/jcb.105.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bordas J., Perez-Grau L., Koch M. H., Vega M. C., Nave C. The superstructure of chromatin and its condensation mechanism. II. Theoretical analysis of the X-ray scattering patterns and model calculations. Eur Biophys J. 1986;13(3):175–185. doi: 10.1007/BF00542561. [DOI] [PubMed] [Google Scholar]
  7. Derenzini M., Farabegoli F. Selective staining of nucleic acids by osmium-ammine complex in thin sections from lowicryl-embedded samples. J Histochem Cytochem. 1990 Oct;38(10):1495–1501. doi: 10.1177/38.10.2205645. [DOI] [PubMed] [Google Scholar]
  8. Felsenfeld G., McGhee J. D. Structure of the 30 nm chromatin fiber. Cell. 1986 Feb 14;44(3):375–377. doi: 10.1016/0092-8674(86)90456-3. [DOI] [PubMed] [Google Scholar]
  9. Fullwood N. J., Meek K. M. A synchroton X-ray study of the changes occurring in the corneal stroma during processing for electron microscopy. J Microsc. 1993 Jan;169(Pt 1):53–60. doi: 10.1111/j.1365-2818.1993.tb03277.x. [DOI] [PubMed] [Google Scholar]
  10. Giannasca P. J., Horowitz R. A., Woodcock C. L. Transitions between in situ and isolated chromatin. J Cell Sci. 1993 Jun;105(Pt 2):551–561. doi: 10.1242/jcs.105.2.551. [DOI] [PubMed] [Google Scholar]
  11. Goldman M. A. The chromatin domain as a unit of gene regulation. Bioessays. 1988 Aug-Sep;9(2-3):50–55. doi: 10.1002/bies.950090204. [DOI] [PubMed] [Google Scholar]
  12. Horowitz R. A., Giannasca P. J., Woodcock C. L. Ultrastructural preservation of nuclei and chromatin: improvement with low-temperature methods. J Microsc. 1990 Feb;157(Pt 2):205–224. doi: 10.1111/j.1365-2818.1990.tb02959.x. [DOI] [PubMed] [Google Scholar]
  13. Horowitz R. A., Woodcock C. L. Alternative staining methods for Lowicryl sections. J Histochem Cytochem. 1992 Jan;40(1):123–133. doi: 10.1177/40.1.1370308. [DOI] [PubMed] [Google Scholar]
  14. Kellum R., Schedl P. A position-effect assay for boundaries of higher order chromosomal domains. Cell. 1991 Mar 8;64(5):941–950. doi: 10.1016/0092-8674(91)90318-s. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lawrence M. C., Jaffer M. A., Sewell B. T. The application of the maximum entropy method to electron microscopic tomography. Ultramicroscopy. 1989 Nov;31(3):285–301. doi: 10.1016/0304-3991(89)90051-x. [DOI] [PubMed] [Google Scholar]
  17. Lohr D., Van Holde K. E. Organization of spacer DNA in chromatin. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6326–6330. doi: 10.1073/pnas.76.12.6326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marko M., Leith A., Parsons D. Three-dimensional reconstruction of cells from serial sections and whole-cell mounts using multilevel contouring of stereo micrographs. J Electron Microsc Tech. 1988 Aug;9(4):395–411. doi: 10.1002/jemt.1060090406. [DOI] [PubMed] [Google Scholar]
  19. Olins A. L., Moyer B. A., Kim S. H., Allison D. P. Synthesis of a more stable osmium ammine electron-dense DNA stain. J Histochem Cytochem. 1989 Mar;37(3):395–398. doi: 10.1177/37.3.2465337. [DOI] [PubMed] [Google Scholar]
  20. Pehrson J. R., Cohen L. H. Effects of DNA looping on pyrimidine dimer formation. Nucleic Acids Res. 1992 Mar 25;20(6):1321–1324. doi: 10.1093/nar/20.6.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pehrson J. R. Thymine dimer formation as a probe of the path of DNA in and between nucleosomes in intact chromatin. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9149–9153. doi: 10.1073/pnas.86.23.9149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rattner J. B., Hamkalo B. A. Higher order structure in metaphase chromosomes. II. The relationship between the 250 A fiber, superbeads and beads-on-a-string. Chromosoma. 1978 Dec 6;69(3):373–379. doi: 10.1007/BF00332140. [DOI] [PubMed] [Google Scholar]
  23. Strauss F., Prunell A. Organization of internucleosomal DNA in rat liver chromatin. EMBO J. 1983;2(1):51–56. doi: 10.1002/j.1460-2075.1983.tb01379.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Subirana J. A., Muñoz-Guerra S., Aymamí J., Radermacher M., Frank J. The layered organization of nucleosomes in 30 nm chromatin fibers. Chromosoma. 1985;91(5):377–390. doi: 10.1007/BF00291012. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Widom J. A relationship between the helical twist of DNA and the ordered positioning of nucleosomes in all eukaryotic cells. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):1095–1099. doi: 10.1073/pnas.89.3.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Widom J. Toward a unified model of chromatin folding. Annu Rev Biophys Biophys Chem. 1989;18:365–395. doi: 10.1146/annurev.bb.18.060189.002053. [DOI] [PubMed] [Google Scholar]
  28. Williams S. P., Athey B. D., Muglia L. J., Schappe R. S., Gough A. H., Langmore J. P. Chromatin fibers are left-handed double helices with diameter and mass per unit length that depend on linker length. Biophys J. 1986 Jan;49(1):233–248. doi: 10.1016/S0006-3495(86)83637-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Woodcock C. L. Chromatin fibers observed in situ in frozen hydrated sections. Native fiber diameter is not correlated with nucleosome repeat length. J Cell Biol. 1994 Apr;125(1):11–19. doi: 10.1083/jcb.125.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Woodcock C. L., Grigoryev S. A., Horowitz R. A., Whitaker N. A chromatin folding model that incorporates linker variability generates fibers resembling the native structures. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):9021–9025. doi: 10.1073/pnas.90.19.9021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Woodcock C. L., McEwen B. F., Frank J. Ultrastructure of chromatin. II. Three-dimensional reconstruction of isolated fibers. J Cell Sci. 1991 May;99(Pt 1):107–114. doi: 10.1242/jcs.99.1.107. [DOI] [PubMed] [Google Scholar]
  33. Woodcock C. L., Woodcock H., Horowitz R. A. Ultrastructure of chromatin. I. Negative staining of isolated fibers. J Cell Sci. 1991 May;99(Pt 1):99–106. doi: 10.1242/jcs.99.1.99. [DOI] [PubMed] [Google Scholar]
  34. Yao J., Lowary P. T., Widom J. Direct detection of linker DNA bending in defined-length oligomers of chromatin. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7603–7607. doi: 10.1073/pnas.87.19.7603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yao J., Lowary P. T., Widom J. Linker DNA bending induced by the core histones of chromatin. Biochemistry. 1991 Aug 27;30(34):8408–8414. doi: 10.1021/bi00098a019. [DOI] [PubMed] [Google Scholar]
  36. 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]

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