Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Jul 1;90(1):222–235. doi: 10.1083/jcb.90.1.222

Structural interaction of cytoskeletal components

M Schliwa, J van Blerkom
PMCID: PMC2111842  PMID: 7019221

Abstract

Three-dimensional cytoskeletal organization of detergent-treated epithelial African green monkey kidney cells (BSC-1) and chick embryo fibroblasts was studied in whole-mount preparations visualized in a high voltage electron microscope. Stereo images are generated at both low and high magnification to reveal both overall cytoskeletal morphology and details of the structural continuity of different filament types. By the use of an improved extraction procedure in combination with heavy meromyosin subfragment 1 decoration of actin filaments, several new features of filament organization are revealed that suggest that the cytoskeleton is a highly interconnected structural unit. In addition to actin filaments, intermediate filaments, and microtubules, a new class of filaments of 2- to 3-nm diameter and 30- to 300-nm length that do not bind heavy merymyosin is demonstrated. They form end-to-side contacts with other cytoskeletal filaments, thereby acting as linkers between various fibers, both like (e.g., actin- actin) and unlike (e.g., actin-intermediate filament, intermediate filament-microtubule). Their nature is unknown. In addition to 2- to 3-nm filaments, actin filaments are demonstrated to form end-to-side contacts with other filaments. Y-shaped actin filament “branches” are observed both in the cell periphery close to ruffles and in more central cell areas also populated by abundant intermediate filaments and microtubules. Arrowhead complexes formed by subfragment 1 decoration of actin filaments point towards the contact site. Actin filaments also form end-to-side contacts with microtubules and intermediate filaments. Careful inspection of numerous actin-microtubule contacts shows that microtubules frequently change their course at sites of contact. A variety of experimentally induced modifications of the frequency of actin-microtubule contacts can be shown to influence the course of microtubules. We conclude that bends in microtubules are imposed by structural interactions with other cytoskeletal elements. A structural and biochemical comparison of whole cells and cytoskeletons demonstrates that the former show a more inticate three-dimensional network and a more complex biochemical composition than the latter. An analysis of the time course of detergent extraction strongly suggests that the cytoskeleton forms a structural backbone with which a large number of proteins of the cytoplasmic ground substance associate in an ordered fashion to form the characteristic image of the “microtrabecular network” (J.J. Wolosewick and K.R. Porter. 1979. J. Cell Biol. 82: 114-139).

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

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

  1. Batten B. E., Aalberg J. J., Anderson E. The cytoplasmic filamentous network in cultured ovarian granulosa cells. Cell. 1980 Oct;21(3):885–895. doi: 10.1016/0092-8674(80)90452-3. [DOI] [PubMed] [Google Scholar]
  2. Ben-Ze'ev A., Duerr A., Solomon F., Penman S. The outer boundary of the cytoskeleton: a lamina derived from plasma membrane proteins. Cell. 1979 Aug;17(4):859–865. doi: 10.1016/0092-8674(79)90326-x. [DOI] [PubMed] [Google Scholar]
  3. Brinkley B. R., Fistel S. H., Marcum J. M., Pardue R. L. Microtubules in cultured cells; indirect immunofluorescent staining with tubulin antibody. Int Rev Cytol. 1980;63:59–95. doi: 10.1016/s0074-7696(08)61757-x. [DOI] [PubMed] [Google Scholar]
  4. Brown S., Levinson W., Spudich J. A. Cytoskeletal elements of chick embryo fibroblasts revealed by detergent extraction. J Supramol Struct. 1976;5(2):119–130. doi: 10.1002/jss.400050203. [DOI] [PubMed] [Google Scholar]
  5. Buckley I. K., Porter K. R. Electron microscopy of critical point dried whole cultured cells. J Microsc. 1975 Jul;104(2):107–120. doi: 10.1111/j.1365-2818.1975.tb04010.x. [DOI] [PubMed] [Google Scholar]
  6. Buckley I. K. Three dimensional fine structure of cultured cells: possible implications for subcellular motility. Tissue Cell. 1975;7(1):51–72. doi: 10.1016/s0040-8166(75)80007-3. [DOI] [PubMed] [Google Scholar]
  7. Fujiwara K., Pollard T. D. Simultaneous localization of myosin and tubulin in human tissue culture cells by double antibody staining. J Cell Biol. 1978 Apr;77(1):182–195. doi: 10.1083/jcb.77.1.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goldman R. D. The role of three cytoplasmic fibers in BHK-21 cell motility. I. Microtubules and the effects of colchicine. J Cell Biol. 1971 Dec;51(3):752–762. doi: 10.1083/jcb.51.3.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gordon W. E., 3rd, Bushnell A., Burridge K. Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum. Cell. 1978 Feb;13(2):249–261. doi: 10.1016/0092-8674(78)90194-0. [DOI] [PubMed] [Google Scholar]
  10. Griffith L. M., Pollard T. D. Evidence for actin filament-microtubule interaction mediated by microtubule-associated proteins. J Cell Biol. 1978 Sep;78(3):958–965. doi: 10.1083/jcb.78.3.958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gröschel-Stewart U. Immunochemistry of cytoplasmic contractile proteins. Int Rev Cytol. 1980;65:193–254. doi: 10.1016/s0074-7696(08)61961-0. [DOI] [PubMed] [Google Scholar]
  12. Hartwig J. H., Tyler J., Stossel T. P. Actin-binding protein promotes the bipolar and perpendicular branching of actin filaments. J Cell Biol. 1980 Dec;87(3 Pt 1):841–848. doi: 10.1083/jcb.87.3.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Henderson D., Weber K. Three-dimensional organization of microfilaments and microtubules in the cytoskeleton. Immunoperoxidase labelling and stereo-electron microscopy of detergent-extracted cells. Exp Cell Res. 1979 Dec;124(2):301–316. doi: 10.1016/0014-4827(79)90206-4. [DOI] [PubMed] [Google Scholar]
  14. Heuser J. E., Kirschner M. W. Filament organization revealed in platinum replicas of freeze-dried cytoskeletons. J Cell Biol. 1980 Jul;86(1):212–234. doi: 10.1083/jcb.86.1.212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ip W., Fischman D. A. High resolution scanning electron microscopy of isolated and in situ cytoskeletal elements. J Cell Biol. 1979 Oct;83(1):249–254. doi: 10.1083/jcb.83.1.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kirschner M. W. Implications of treadmilling for the stability and polarity of actin and tubulin polymers in vivo. J Cell Biol. 1980 Jul;86(1):330–334. doi: 10.1083/jcb.86.1.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lenk R., Ransom L., Kaufmann Y., Penman S. A cytoskeletal structure with associated polyribosomes obtained from HeLa cells. Cell. 1977 Jan;10(1):67–78. doi: 10.1016/0092-8674(77)90141-6. [DOI] [PubMed] [Google Scholar]
  18. Maruyama K., Kaibara M., Fukada E. Rheology of F-actin. I. Network of F-actin in solution. Biochim Biophys Acta. 1974 Nov 5;371(1):20–29. doi: 10.1016/0005-2795(74)90150-0. [DOI] [PubMed] [Google Scholar]
  19. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  20. Osborn M., Born T., Koitsch H. J., Weber K. Stereo immunofluorescence microscopy: I. Three-dimensional arrangement of microfilaments, microtubules and tonofilaments. Cell. 1978 Jul;14(3):477–488. doi: 10.1016/0092-8674(78)90234-9. [DOI] [PubMed] [Google Scholar]
  21. Osborn M., Weber K. The detertent-resistant cytoskeleton of tissue culture cells includes the nucleus and the microfilament bundles. Exp Cell Res. 1977 May;106(2):339–349. doi: 10.1016/0014-4827(77)90179-3. [DOI] [PubMed] [Google Scholar]
  22. Pudney J., Singer R. H. Electron microscopic visualization of the filamentous reticulum in whole cultured presumptive chick myoblasts. Am J Anat. 1979 Nov;156(3):321–336. doi: 10.1002/aja.1001560304. [DOI] [PubMed] [Google Scholar]
  23. Schiff P. B., Fant J., Horwitz S. B. Promotion of microtubule assembly in vitro by taxol. Nature. 1979 Feb 22;277(5698):665–667. doi: 10.1038/277665a0. [DOI] [PubMed] [Google Scholar]
  24. Schliwa M., Euteneuer U., Bulinski J. C., Izant J. G. Calcium lability of cytoplasmic microtubules and its modulation by microtubule-associated proteins. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1037–1041. doi: 10.1073/pnas.78.2.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shelanski M. L., Gaskin F., Cantor C. R. Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci U S A. 1973 Mar;70(3):765–768. doi: 10.1073/pnas.70.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Small J. V., Celis J. E. Direct visualization of the 10-nm (100-A)-filament network in whole and enucleated cultured cells. J Cell Sci. 1978 Jun;31:393–409. doi: 10.1242/jcs.31.1.393. [DOI] [PubMed] [Google Scholar]
  27. Small J. V., Sobieszek A. Studies on the function and composition of the 10-NM(100-A) filaments of vertebrate smooth muscle. J Cell Sci. 1977 Feb;23:243–268. doi: 10.1242/jcs.23.1.243. [DOI] [PubMed] [Google Scholar]
  28. Summers K., Kirschner M. W. Characteristics of the polar assembly and disassembly of microtubules observed in vitro by darkfield light microscopy. J Cell Biol. 1979 Oct;83(1):205–217. doi: 10.1083/jcb.83.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Telzer B. R., Rosenbaum J. L. Cell cycle-dependent, in vitro assembly of microtubules onto pericentriolar material of HeLa cells. J Cell Biol. 1979 Jun;81(3):484–497. doi: 10.1083/jcb.81.3.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Temmink J. H., Spiele H. Preservation of cytoskeletal elements for electron microscopy. Cell Biol Int Rep. 1978 Jan;2(1):51–59. doi: 10.1016/0309-1651(78)90084-x. [DOI] [PubMed] [Google Scholar]
  31. Tilney L. G., Derosier D. J., Mulroy M. J. The organization of actin filaments in the stereocilia of cochlear hair cells. J Cell Biol. 1980 Jul;86(1):244–259. doi: 10.1083/jcb.86.1.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Trotter J. A., Foerder B. A., Keller J. M. Intracellular fibres in cultured cells: analysis by scanning and transmission electron microscopy and by SDS-polyacrylamide gel electrophoresis. J Cell Sci. 1978 Jun;31:369–392. doi: 10.1242/jcs.31.1.369. [DOI] [PubMed] [Google Scholar]
  33. Weber K., Rathke P. C., Osborn M. Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1820–1824. doi: 10.1073/pnas.75.4.1820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Webster R. E., Henderson D., Osborn M., Weber K. Three-dimensional electron microscopical visualization of the cytoskeleton of animal cells: immunoferritin identification of actin- and tubulin-containing structures. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5511–5515. doi: 10.1073/pnas.75.11.5511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Webster R. E., Osborn M., Weber K. Visualization of the same PtK2 cytoskeletons by both immunofluorescence and low power electron microscopy. Exp Cell Res. 1978 Nov;117(1):47–61. doi: 10.1016/0014-4827(78)90426-3. [DOI] [PubMed] [Google Scholar]
  36. Wieland T. Modification of actins by phallotoxins. Naturwissenschaften. 1977 Jun;64(6):303–309. doi: 10.1007/BF00446784. [DOI] [PubMed] [Google Scholar]
  37. Wolosewick J. J., Porter K. R. Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality. J Cell Biol. 1979 Jul;82(1):114–139. doi: 10.1083/jcb.82.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wolosewick J. J., Porter K. R. Stereo high-voltage electron microscopy of whole cells of the human diploid line, WI-38. Am J Anat. 1976 Nov;147(3):303–323. doi: 10.1002/aja.1001470305. [DOI] [PubMed] [Google Scholar]
  39. Woodrum D. T., Rich S. A., Pollard T. D. Evidence for biased bidirectional polymerization of actin filaments using heavy meromyosin prepared by an improved method. J Cell Biol. 1975 Oct;67(1):231–237. doi: 10.1083/jcb.67.1.231. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES