Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Dec 1;97(6):1788–1794. doi: 10.1083/jcb.97.6.1788

Rearrangement of the keratin cytoskeleton after combined treatment with microtubule and microfilament inhibitors

PMCID: PMC2112713  PMID: 6196368

Abstract

In addition to containing microtubule and microfilament systems, vertebrate epithelial cells contain an elaborate keratin intermediate- filament cytoskeleton. Little is known about its structural organization or function. Using indirect immunofluorescence microscopy with an antikeratin antiserum probe, we found that destabilization of microtubules and microfilaments with cytostatic drugs induces significant alterations in the cytoskeletal organization of keratin filaments in HeLa and fetal mouse epidermal cells. Keratin filament organization was observed to undergo a rapid (1-2 h) transition from a uniform distribution to an open lattice of keratin fibers stabilized by membrane-associated focal centers. Since addition of any one drug alone did not elicit significant organizational change in the keratin cytoskeleton, we suggest that microfilaments and microtubules have a combined role in maintaining the arrangement of keratin in these cells. Vimentin filaments, the only other intermediate-sized filaments found in HeLa cells, did not co-distribute with keratin in untreated or drug- treated cells. These findings offer a new way to approach the study of the dynamics and functional roles of the keratin cytoskeleton in epithelial cells.

Full Text

The Full Text of this article is available as a PDF (878.1 KB).

Selected References

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

  1. Eckert B. S., Daley R. A., Parysek L. M. In vivo disruption of the cytokeratin cytoskeleton in cultured epithelial cells by microinjection of antikeratin: evidence for the presence of an intermediate-filament-organizing center. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):403–412. doi: 10.1101/sqb.1982.046.01.039. [DOI] [PubMed] [Google Scholar]
  2. Franke W. W., Schmid E., Grund C., Geiger B. Intermediate filament proteins in nonfilamentous structures: transient disintegration and inclusion of subunit proteins in granular aggregates. Cell. 1982 Aug;30(1):103–113. doi: 10.1016/0092-8674(82)90016-2. [DOI] [PubMed] [Google Scholar]
  3. Franke W. W., Schmid E., Grund C., Müller H., Engelbrecht I., Moll R., Stadler J., Jarasch E. D. Antibodies to high molecular weight polypeptides of desmosomes: specific localization of a class of junctional proteins in cells and tissue. Differentiation. 1981;20(3):217–241. doi: 10.1111/j.1432-0436.1981.tb01178.x. [DOI] [PubMed] [Google Scholar]
  4. Franke W. W., Schmid E., Osborn M., Weber K. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5034–5038. doi: 10.1073/pnas.75.10.5034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Franke W. W., Schmid E., Weber K., Osborn M. HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res. 1979 Jan;118(1):95–109. doi: 10.1016/0014-4827(79)90587-1. [DOI] [PubMed] [Google Scholar]
  6. Franke W. W., Weber K., Osborn M., Schmid E., Freudenstein C. Antibody to prekeratin. Decoration of tonofilament like arrays in various cells of epithelial character. Exp Cell Res. 1978 Oct 15;116(2):429–445. doi: 10.1016/0014-4827(78)90466-4. [DOI] [PubMed] [Google Scholar]
  7. Geiger B., Schmid E., Franke W. W. Spatial distribution of proteins specific for desmosomes and adhaerens junctions in epithelial cells demonstrated by double immunofluorescence microscopy. Differentiation. 1983;23(3):189–205. doi: 10.1111/j.1432-0436.1982.tb01283.x. [DOI] [PubMed] [Google Scholar]
  8. Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  9. Horwitz B., Kupfer H., Eshhar Z., Geiger B. Reorganization of arrays of prekeratin filaments during mitosis. Immunofluorescence microscopy with multiclonal and monoclonal prekeratin antibodies. Exp Cell Res. 1981 Aug;134(2):281–290. doi: 10.1016/0014-4827(81)90427-4. [DOI] [PubMed] [Google Scholar]
  10. Kelly D. E. Fine structure of desmosomes. , hemidesmosomes, and an adepidermal globular layer in developing newt epidermis. J Cell Biol. 1966 Jan;28(1):51–72. doi: 10.1083/jcb.28.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Keski-Oja J., Lehto V. P., Virtanen I. Keratin filaments of mouse epithelial cells are rapidly affected by epidermal growth factor. J Cell Biol. 1981 Aug;90(2):537–541. doi: 10.1083/jcb.90.2.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klymkowsky M. W., Miller R. H., Lane E. B. Morphology, behavior, and interaction of cultured epithelial cells after the antibody-induced disruption of keratin filament organization. J Cell Biol. 1983 Feb;96(2):494–509. doi: 10.1083/jcb.96.2.494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knapp L. W., O'Guin W. M., Sawyer R. H. Drug-induced alterations of cytokeratin organization in cultured epithelial cells. Science. 1983 Feb 4;219(4584):501–503. doi: 10.1126/science.6186022. [DOI] [PubMed] [Google Scholar]
  14. Lazarides E. Actin, alpha-actinin, and tropomyosin interaction in the structural organization of actin filaments in nonmuscle cells. J Cell Biol. 1976 Feb;68(2):202–219. doi: 10.1083/jcb.68.2.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  16. Lazarides E. Intermediate filaments: a chemically heterogeneous, developmentally regulated class of proteins. Annu Rev Biochem. 1982;51:219–250. doi: 10.1146/annurev.bi.51.070182.001251. [DOI] [PubMed] [Google Scholar]
  17. O'Guin W. M., Sawyer R. H. Avian scale development. VII. Relationships between morphogenetic and biosynthetic differentiation. Dev Biol. 1982 Feb;89(2):485–492. doi: 10.1016/0012-1606(82)90336-0. [DOI] [PubMed] [Google Scholar]
  18. Overton J. Experiments with junctions of the adhaerens type. Curr Top Dev Biol. 1975;10:1–34. doi: 10.1016/s0070-2153(08)60037-x. [DOI] [PubMed] [Google Scholar]
  19. Paulin D., Babinet C., Weber K., Osborn M. Antibodies as probes of cellular differentiation and cytoskeletal organization in the mouse blastocyst. Exp Cell Res. 1980 Dec;130(2):297–304. doi: 10.1016/0014-4827(80)90006-3. [DOI] [PubMed] [Google Scholar]
  20. Skerrow C. J., Matoltsy A. G. Chemical characterization of isolated epidermal desmosomes. J Cell Biol. 1974 Nov;63(2 Pt 1):524–530. doi: 10.1083/jcb.63.2.524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Staehelin L. A. Structure and function of intercellular junctions. Int Rev Cytol. 1974;39:191–283. doi: 10.1016/s0074-7696(08)60940-7. [DOI] [PubMed] [Google Scholar]
  22. Sun T. T., Green H. Immunofluorescent staining of keratin fibers in cultured cells. Cell. 1978 Jul;14(3):469–476. doi: 10.1016/0092-8674(78)90233-7. [DOI] [PubMed] [Google Scholar]
  23. Sun T. T., Shih C., Green H. Keratin cytoskeletons in epithelial cells of internal organs. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2813–2817. doi: 10.1073/pnas.76.6.2813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wessells N. K., Spooner B. S., Ash J. F., Bradley M. O., Luduena M. A., Taylor E. L., Wrenn J. T., Yamada K. Microfilaments in cellular and developmental processes. Science. 1971 Jan 15;171(3967):135–143. doi: 10.1126/science.171.3967.135. [DOI] [PubMed] [Google Scholar]

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

RESOURCES