Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Jul 1;99(1 Pt 2):83s–91s. doi: 10.1083/jcb.99.1.83s

Microfilament-organizing centers in areas of cell contact: cytoskeletal interactions during cell attachment and locomotion

B Geiger, Z Avnur, G Rinnerthaler, H Hinssen, V J Small
PMCID: PMC2275602  PMID: 6430912

Abstract

In this article we discuss three aspects of cell contact formation: (a) the molecular architecture of the cytomatrix in cell-to-substrate focal contacts, (b) the dynamic properties of membrane- and microfilament-associated proteins in the contact areas, and (c) the involvement of microtubules in the coordinated and directed formation of new substrate contacts during cell locomotion. We show that different microfilament-associated proteins exhibit distinct patterns of association with focal contacts: some proteins are specifically associated with focal contacts (vinculin and talin); alpha-actinin is enriched in the contact areas but also is present along the stress fibers and in the lamellipodium; actin and filamin are detected throughout the contact areas but in apparently reduced amounts compared with the associated stress fibers; and tropomyosin, myosin, and spectrin are either absent from the endofacial surfaces of contact areas or are present in only very small amounts. Fluorescence photobleaching recovery analyses performed with living cells microinjected with fluorescently labeled actin, vinculin, and alpha-actinin indicate that each of these proteins maintains a dynamic equilibrium between a soluble cytoplasmic pool and a membrane-bound fraction. Correlation of the distribution of vinculin and tubulin in motile fibroblasts to local movements of the leading edge of the same cells indicates that free-end microtubules extend into actively ruffling areas along the lamellipodium and that new vinculin-containing contacts are preferentially formed in these protruding regions.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Abercrombie M., Dunn G. A. Adhesions of fibroblasts to substratum during contact inhibition observed by interference reflection microscopy. Exp Cell Res. 1975 Apr;92(1):57–62. doi: 10.1016/0014-4827(75)90636-9. [DOI] [PubMed] [Google Scholar]
  2. Abercrombie M., Heaysman J. E., Pegrum S. M. The locomotion of fibroblasts in culture. I. Movements of the leading edge. Exp Cell Res. 1970 Mar;59(3):393–398. doi: 10.1016/0014-4827(70)90646-4. [DOI] [PubMed] [Google Scholar]
  3. Abercrombie M., Heaysman J. E., Pegrum S. M. The locomotion of fibroblasts in culture. IV. Electron microscopy of the leading lamella. Exp Cell Res. 1971 Aug;67(2):359–367. doi: 10.1016/0014-4827(71)90420-4. [DOI] [PubMed] [Google Scholar]
  4. Albrecht-Buehler G., Bushnell A. The orientation of centrioles in migrating 3T3 cells. Exp Cell Res. 1979 Apr;120(1):111–118. doi: 10.1016/0014-4827(79)90542-1. [DOI] [PubMed] [Google Scholar]
  5. Albrecht-Buehler G. Does the geometric design of centrioles imply their function? Cell Motil. 1981;1(2):237–245. doi: 10.1002/cm.970010206. [DOI] [PubMed] [Google Scholar]
  6. Avnur Z., Geiger B. Substrate-attached membranes of cultured cells isolation and characterization of ventral cell membranes and the associated cytoskeleton. J Mol Biol. 1981 Dec 5;153(2):361–379. doi: 10.1016/0022-2836(81)90283-7. [DOI] [PubMed] [Google Scholar]
  7. Avnur Z., Small J. V., Geiger B. Actin-independent association of vinculin with the cytoplasmic aspect of the plasma membrane in cell-contact areas. J Cell Biol. 1983 Jun;96(6):1622–1630. doi: 10.1083/jcb.96.6.1622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burridge K., Connell L. A new protein of adhesion plaques and ruffling membranes. J Cell Biol. 1983 Aug;97(2):359–367. doi: 10.1083/jcb.97.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Burridge K., Kelly T., Mangeat P. Nonerythrocyte spectrins: actin-membrane attachment proteins occurring in many cell types. J Cell Biol. 1982 Nov;95(2 Pt 1):478–486. doi: 10.1083/jcb.95.2.478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chen W. T. Mechanism of retraction of the trailing edge during fibroblast movement. J Cell Biol. 1981 Jul;90(1):187–200. doi: 10.1083/jcb.90.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Brabander M., De Mey J., Van de Veire R., Aerts F., Geuens G. Microtubules in mammalian cell shape and surface modulation: an alternative hypothesis. Cell Biol Int Rep. 1977 Sep;1(5):453–461. doi: 10.1016/0309-1651(77)90080-7. [DOI] [PubMed] [Google Scholar]
  12. Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. doi: 10.1016/0092-8674(79)90368-4. [DOI] [PubMed] [Google Scholar]
  13. Geiger B., Avnur Z., Kreis T. E., Schlessinger J. The dynamics of cytoskeletal organization in areas of cell contact. Cell Muscle Motil. 1984;5:195–234. doi: 10.1007/978-1-4684-4592-3_5. [DOI] [PubMed] [Google Scholar]
  14. Geiger B., Avnur Z., Schlessinger J. Restricted mobility of membrane constituents in cell-substrate focal contacts of chicken fibroblasts. J Cell Biol. 1982 May;93(2):495–500. doi: 10.1083/jcb.93.2.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Geiger B., Dutton A. H., Tokuyasu K. T., Singer S. J. Immunoelectron microscope studies of membrane-microfilament interactions: distributions of alpha-actinin, tropomyosin, and vinculin in intestinal epithelial brush border and chicken gizzard smooth muscle cells. J Cell Biol. 1981 Dec;91(3 Pt 1):614–628. doi: 10.1083/jcb.91.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Geiger B. Involvement of vinculin in contact-induced cytoskeletal interactions. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):671–682. doi: 10.1101/sqb.1982.046.01.063. [DOI] [PubMed] [Google Scholar]
  17. Geiger B., Singer S. J. Association of microtubules and intermediate filaments in chicken gizzard cells as detected by double immunofluorescence. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4769–4773. doi: 10.1073/pnas.77.8.4769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Geiger B., Singer S. J. The participation of alpha-actinin in the capping of cell membrane components. Cell. 1979 Jan;16(1):213–222. doi: 10.1016/0092-8674(79)90202-2. [DOI] [PubMed] [Google Scholar]
  19. Geiger B. The association of rhodamine - labelled alpha-actinin with actin bundles in demembranated cells. Cell Biol Int Rep. 1981 Jun;5(6):627–634. doi: 10.1016/s0309-1651(81)80015-x. [DOI] [PubMed] [Google Scholar]
  20. Geiger B., Tokuyasu K. T., Dutton A. H., Singer S. J. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. doi: 10.1073/pnas.77.7.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gingell D. The interpretation of interference-reflection images of spread cells: significant contributions from thin peripheral cytoplasm. J Cell Sci. 1981 Jun;49:237–247. doi: 10.1242/jcs.49.1.237. [DOI] [PubMed] [Google Scholar]
  22. Heath J. P., Dunn G. A. Cell to substratum contacts of chick fibroblasts and their relation to the microfilament system. A correlated interference-reflexion and high-voltage electron-microscope study. J Cell Sci. 1978 Feb;29:197–212. doi: 10.1242/jcs.29.1.197. [DOI] [PubMed] [Google Scholar]
  23. Hinssen H. An actin-modulating protein from Physarum polycephalum. I. Isolation and purification. Eur J Cell Biol. 1981 Feb;23(2):225–233. [PubMed] [Google Scholar]
  24. Hinssen H. An actin-modulating protein from Physarum polycephalum. II. Ca++-dependence and other properties. Eur J Cell Biol. 1981 Feb;23(2):234–240. [PubMed] [Google Scholar]
  25. Izzard C. S., Lochner L. R. Cell-to-substrate contacts in living fibroblasts: an interference reflexion study with an evaluation of the technique. J Cell Sci. 1976 Jun;21(1):129–159. doi: 10.1242/jcs.21.1.129. [DOI] [PubMed] [Google Scholar]
  26. Izzard C. S., Lochner L. R. Formation of cell-to-substrate contacts during fibroblast motility: an interference-reflexion study. J Cell Sci. 1980 Apr;42:81–116. doi: 10.1242/jcs.42.1.81. [DOI] [PubMed] [Google Scholar]
  27. Jockusch B. M., Isenberg G. Interaction of alpha-actinin and vinculin with actin: opposite effects on filament network formation. Proc Natl Acad Sci U S A. 1981 May;78(5):3005–3009. doi: 10.1073/pnas.78.5.3005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kreis T. E., Geiger B., Schlessinger J. Mobility of microinjected rhodamine actin within living chicken gizzard cells determined by fluorescence photobleaching recovery. Cell. 1982 Jul;29(3):835–845. doi: 10.1016/0092-8674(82)90445-7. [DOI] [PubMed] [Google Scholar]
  29. Lazarides E., Burridge K. Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells. Cell. 1975 Nov;6(3):289–298. doi: 10.1016/0092-8674(75)90180-4. [DOI] [PubMed] [Google Scholar]
  30. Lin S., Wilkins J. A., Cribbs D. H., Grumet M., Lin D. C. Proteins and complexes that affect actin-filament assembly and interactions. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):625–632. doi: 10.1101/sqb.1982.046.01.058. [DOI] [PubMed] [Google Scholar]
  31. Lindberg U., Carlsson L., Markey F., Nyström L. E. The unpolymerised form of actin in non-muscle cells. Methods Achiev Exp Pathol. 1979;8:143–170. [PubMed] [Google Scholar]
  32. Otto J. J. Detection of vinculin-binding proteins with an 125I-vinculin gel overlay technique. J Cell Biol. 1983 Oct;97(4):1283–1287. doi: 10.1083/jcb.97.4.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schlessinger J., Axelrod D., Koppel D. E., Webb W. W., Elson E. L. Lateral transport of a lipid probe and labeled proteins on a cell membrane. Science. 1977 Jan 21;195(4275):307–309. doi: 10.1126/science.556653. [DOI] [PubMed] [Google Scholar]
  34. Schlessinger J., Koppel D. E., Axelrod D., Jacobson K., Webb W. W., Elson E. L. Lateral transport on cell membranes: mobility of concanavalin A receptors on myoblasts. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2409–2413. doi: 10.1073/pnas.73.7.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Singer S. J., Ball E. H., Geiger B., Chen W. T. Immunolabeling studies of cytoskeletal association in cultured cells. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):303–316. doi: 10.1101/sqb.1982.046.01.032. [DOI] [PubMed] [Google Scholar]
  36. Small J. V. Organization of actin in the leading edge of cultured cells: influence of osmium tetroxide and dehydration on the ultrastructure of actin meshworks. J Cell Biol. 1981 Dec;91(3 Pt 1):695–705. doi: 10.1083/jcb.91.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Small J. V., Rinnerthaler G., Hinssen H. Organization of actin meshworks in cultured cells: the leading edge. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):599–611. doi: 10.1101/sqb.1982.046.01.056. [DOI] [PubMed] [Google Scholar]
  38. Tokuyasu K. T., Dutton A. H., Geiger B., Singer S. J. Ultrastructure of chicken cardiac muscle as studied by double immunolabeling in electron microscopy. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7619–7623. doi: 10.1073/pnas.78.12.7619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vasiliev J. M. Spreading and locomotion of tissue cells: factors controlling the distribution of pseudopodia. Philos Trans R Soc Lond B Biol Sci. 1982 Nov 4;299(1095):159–167. doi: 10.1098/rstb.1982.0122. [DOI] [PubMed] [Google Scholar]

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

RESOURCES