Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Mar 2;120(6):1381–1391. doi: 10.1083/jcb.120.6.1381

Relative distribution of actin, myosin I, and myosin II during the wound healing response of fibroblasts

PMCID: PMC2119744  PMID: 8449984

Abstract

Myosin I is present in Swiss 3T3 fibroblasts and its localization reflects a possible involvement in the extension and/or retraction of protrusions at the leading edge of locomoting cells and the transport of vesicles, but not in the contraction of stress fibers or transverse fibers. An affinity-purified polyclonal antibody to brush border myosin I colocalizes with a polypeptide of 120 kD in fibroblast extracts. Within initial protrusions of polarized, migrating fibroblasts, myosin I exhibits a punctate distribution, whereas actin is diffuse and myosin II is absent. Myosin I also exists in linear arrays parallel to the direction of migration in filopodia and microspikes, established protrusions, and within the leading lamellae of migrating cells. Myosin II and actin colocalize along transverse fibers in the lamellae of migrating cells, while myosin I displays no definitive organization along these fibers. During contractions of actin-based fibers, myosin II is concentrated in the center of the cell, while the distribution of myosin I does not change. Thus, myosin I is found at the correct location and time to be involved in the extension and/or retraction of protrusions and the transport of vesicles. Myosin II-based contractions in more posterior cellular regions could generate forces to separate cells, maintain a polarized cell shape, maintain the direction of locomotion, maximize the rate of locomotion, and/or aid in the delivery of cytoskeletal/contractile subunits to the leading edge.

Full Text

The Full Text of this article is available as a PDF (6.5 MB).

Selected References

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

  1. Adams R. J., Pollard T. D. Binding of myosin I to membrane lipids. Nature. 1989 Aug 17;340(6234):565–568. doi: 10.1038/340565a0. [DOI] [PubMed] [Google Scholar]
  2. Adams R. J., Pollard T. D. Propulsion of organelles isolated from Acanthamoeba along actin filaments by myosin-I. Nature. 1986 Aug 21;322(6081):754–756. doi: 10.1038/322754a0. [DOI] [PubMed] [Google Scholar]
  3. Barylko B., Wagner M. C., Reizes O., Albanesi J. P. Purification and characterization of a mammalian myosin I. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):490–494. doi: 10.1073/pnas.89.2.490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bray D., White J. G. Cortical flow in animal cells. Science. 1988 Feb 19;239(4842):883–888. doi: 10.1126/science.3277283. [DOI] [PubMed] [Google Scholar]
  5. Brundage R. A., Fogarty K. E., Tuft R. A., Fay F. S. Calcium gradients underlying polarization and chemotaxis of eosinophils. Science. 1991 Nov 1;254(5032):703–706. doi: 10.1126/science.1948048. [DOI] [PubMed] [Google Scholar]
  6. Cao L. G., Wang Y. L. Mechanism of the formation of contractile ring in dividing cultured animal cells. I. Recruitment of preexisting actin filaments into the cleavage furrow. J Cell Biol. 1990 Apr;110(4):1089–1095. doi: 10.1083/jcb.110.4.1089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Collins K., Matsudaira P. Differential regulation of vertebrate myosins I and II. J Cell Sci Suppl. 1991;14:11–16. doi: 10.1242/jcs.1991.supplement_14.3. [DOI] [PubMed] [Google Scholar]
  8. Collins K., Sellers J. R., Matsudaira P. Calmodulin dissociation regulates brush border myosin I (110-kD-calmodulin) mechanochemical activity in vitro. J Cell Biol. 1990 Apr;110(4):1137–1147. doi: 10.1083/jcb.110.4.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Coluccio L. M. Identification of the microvillar 110-kDa calmodulin complex (myosin-1) in kidney. Eur J Cell Biol. 1991 Dec;56(2):286–294. [PubMed] [Google Scholar]
  10. Condeelis J. Are all pseudopods created equal? Cell Motil Cytoskeleton. 1992;22(1):1–6. doi: 10.1002/cm.970220102. [DOI] [PubMed] [Google Scholar]
  11. Condeelis J., Bresnick A., Demma M., Dharmawardhane S., Eddy R., Hall A. L., Sauterer R., Warren V. Mechanisms of amoeboid chemotaxis: an evaluation of the cortical expansion model. Dev Genet. 1990;11(5-6):333–340. doi: 10.1002/dvg.1020110504. [DOI] [PubMed] [Google Scholar]
  12. Conrad P. A., Nederlof M. A., Herman I. M., Taylor D. L. Correlated distribution of actin, myosin, and microtubules at the leading edge of migrating Swiss 3T3 fibroblasts. Cell Motil Cytoskeleton. 1989;14(4):527–543. doi: 10.1002/cm.970140410. [DOI] [PubMed] [Google Scholar]
  13. De Lozanne A., Spudich J. A. Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. Science. 1987 May 29;236(4805):1086–1091. doi: 10.1126/science.3576222. [DOI] [PubMed] [Google Scholar]
  14. DeBiasio R. L., Wang L. L., Fisher G. W., Taylor D. L. The dynamic distribution of fluorescent analogues of actin and myosin in protrusions at the leading edge of migrating Swiss 3T3 fibroblasts. J Cell Biol. 1988 Dec;107(6 Pt 2):2631–2645. doi: 10.1083/jcb.107.6.2631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Drenckhahn D., Dermietzel R. Organization of the actin filament cytoskeleton in the intestinal brush border: a quantitative and qualitative immunoelectron microscope study. J Cell Biol. 1988 Sep;107(3):1037–1048. doi: 10.1083/jcb.107.3.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ezzell R. M., Leung J., Collins K., Chafel M. M., Cardozo T. J., Matsudaira P. T. Expression and localization of villin, fimbrin, and myosin I in differentiating mouse F9 teratocarcinoma cells. Dev Biol. 1992 Jun;151(2):575–585. doi: 10.1016/0012-1606(92)90195-m. [DOI] [PubMed] [Google Scholar]
  17. Fisher G. W., Conrad P. A., DeBiasio R. L., Taylor D. L. Centripetal transport of cytoplasm, actin, and the cell surface in lamellipodia of fibroblasts. Cell Motil Cytoskeleton. 1988;11(4):235–247. doi: 10.1002/cm.970110403. [DOI] [PubMed] [Google Scholar]
  18. Fujisaki H., Albanesi J. P., Korn E. D. Experimental evidence for the contractile activities of Acanthamoeba myosins IA and IB. J Biol Chem. 1985 Sep 15;260(20):11183–11189. [PubMed] [Google Scholar]
  19. Fukui Y., Inoué S. Cell division in Dictyostelium with special emphasis on actomyosin organization in cytokinesis. Cell Motil Cytoskeleton. 1991;18(1):41–54. doi: 10.1002/cm.970180105. [DOI] [PubMed] [Google Scholar]
  20. Fukui Y., Lynch T. J., Brzeska H., Korn E. D. Myosin I is located at the leading edges of locomoting Dictyostelium amoebae. Nature. 1989 Sep 28;341(6240):328–331. doi: 10.1038/341328a0. [DOI] [PubMed] [Google Scholar]
  21. Gadasi H., Korn E. D. Evidence for differential intracellular localization of the Acanthamoeba myosin isoenzymes. Nature. 1980 Jul 31;286(5772):452–456. doi: 10.1038/286452a0. [DOI] [PubMed] [Google Scholar]
  22. Giuliano K. A., Kolega J., DeBiasio R. L., Taylor D. L. Myosin II phosphorylation and the dynamics of stress fibers in serum-deprived and stimulated fibroblasts. Mol Biol Cell. 1992 Sep;3(9):1037–1048. doi: 10.1091/mbc.3.9.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Giuliano K. A., Taylor D. L. Formation, transport, contraction, and disassembly of stress fibers in fibroblasts. Cell Motil Cytoskeleton. 1990;16(1):14–21. doi: 10.1002/cm.970160104. [DOI] [PubMed] [Google Scholar]
  24. Hahn K., DeBiasio R., Taylor D. L. Patterns of elevated free calcium and calmodulin activation in living cells. Nature. 1992 Oct 22;359(6397):736–738. doi: 10.1038/359736a0. [DOI] [PubMed] [Google Scholar]
  25. Halsall D. J., Hammer J. A., 3rd A second isoform of chicken brush border myosin I contains a 29-residue inserted sequence that binds calmodulin. FEBS Lett. 1990 Jul 2;267(1):126–130. doi: 10.1016/0014-5793(90)80305-3. [DOI] [PubMed] [Google Scholar]
  26. Hammer J. A., 3rd, Bowers B., Paterson B. M., Korn E. D. Complete nucleotide sequence and deduced polypeptide sequence of a nonmuscle myosin heavy chain gene from Acanthamoeba: evidence of a hinge in the rodlike tail. J Cell Biol. 1987 Aug;105(2):913–925. doi: 10.1083/jcb.105.2.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hammer J. A., 3rd, Sellers J. R., Korn E. D. Phosphorylation and activation of smooth muscle myosin by Acanthamoeba myosin I heavy chain kinase. J Biol Chem. 1984 Mar 10;259(5):3224–3229. [PubMed] [Google Scholar]
  28. Hayden S. M., Wolenski J. S., Mooseker M. S. Binding of brush border myosin I to phospholipid vesicles. J Cell Biol. 1990 Aug;111(2):443–451. doi: 10.1083/jcb.111.2.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Höner B., Citi S., Kendrick-Jones J., Jockusch B. M. Modulation of cellular morphology and locomotory activity by antibodies against myosin. J Cell Biol. 1988 Dec;107(6 Pt 1):2181–2189. doi: 10.1083/jcb.107.6.2181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Janson L. W., Kolega J., Taylor D. L. Modulation of contraction by gelation/solation in a reconstituted motile model. J Cell Biol. 1991 Sep;114(5):1005–1015. doi: 10.1083/jcb.114.5.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Jung G., Hammer J. A., 3rd Generation and characterization of Dictyostelium cells deficient in a myosin I heavy chain isoform. J Cell Biol. 1990 Jun;110(6):1955–1964. doi: 10.1083/jcb.110.6.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Jung G., Saxe C. L., 3rd, Kimmel A. R., Hammer J. A., 3rd Dictyostelium discoideum contains a gene encoding a myosin I heavy chain. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6186–6190. doi: 10.1073/pnas.86.16.6186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Jung G., Schmidt C. J., Hammer J. A., 3rd Myosin I heavy-chain genes of Acanthamoeba castellanii: cloning of a second gene and evidence for the existence of a third isoform. Gene. 1989 Oct 30;82(2):269–280. doi: 10.1016/0378-1119(89)90052-8. [DOI] [PubMed] [Google Scholar]
  34. Kamm K. E., Stull J. T. Regulation of smooth muscle contractile elements by second messengers. Annu Rev Physiol. 1989;51:299–313. doi: 10.1146/annurev.ph.51.030189.001503. [DOI] [PubMed] [Google Scholar]
  35. Kolega J., Janson L. W., Taylor D. L. The role of solation-contraction coupling in regulating stress fiber dynamics in nonmuscle cells. J Cell Biol. 1991 Sep;114(5):993–1003. doi: 10.1083/jcb.114.5.993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Korn E. D., Hammer J. A., 3rd Myosin I. Curr Opin Cell Biol. 1990 Feb;2(1):57–61. doi: 10.1016/s0955-0674(05)80031-6. [DOI] [PubMed] [Google Scholar]
  37. Manstein D. J., Titus M. A., De Lozanne A., Spudich J. A. Gene replacement in Dictyostelium: generation of myosin null mutants. EMBO J. 1989 Mar;8(3):923–932. doi: 10.1002/j.1460-2075.1989.tb03453.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. McNeil P. L., McKenna M. P., Taylor D. L. A transient rise in cytosolic calcium follows stimulation of quiescent cells with growth factors and is inhibitable with phorbol myristate acetate. J Cell Biol. 1985 Aug;101(2):372–379. doi: 10.1083/jcb.101.2.372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Melan M. A., Sluder G. Redistribution and differential extraction of soluble proteins in permeabilized cultured cells. Implications for immunofluorescence microscopy. J Cell Sci. 1992 Apr;101(Pt 4):731–743. doi: 10.1242/jcs.101.4.731. [DOI] [PubMed] [Google Scholar]
  40. Miyata H., Bowers B., Korn E. D. Plasma membrane association of Acanthamoeba myosin I. J Cell Biol. 1989 Oct;109(4 Pt 1):1519–1528. doi: 10.1083/jcb.109.4.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Mooseker M. S., Conzelman K. A., Coleman T. R., Heuser J. E., Sheetz M. P. Characterization of intestinal microvillar membrane disks: detergent-resistant membrane sheets enriched in associated brush border myosin I (110K-calmodulin). J Cell Biol. 1989 Sep;109(3):1153–1161. doi: 10.1083/jcb.109.3.1153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Oster G. F., Perelson A. S. The physics of cell motility. J Cell Sci Suppl. 1987;8:35–54. doi: 10.1242/jcs.1987.supplement_8.3. [DOI] [PubMed] [Google Scholar]
  43. Pollard T. D., Doberstein S. K., Zot H. G. Myosin-I. Annu Rev Physiol. 1991;53:653–681. doi: 10.1146/annurev.ph.53.030191.003253. [DOI] [PubMed] [Google Scholar]
  44. Pollard T. D., Satterwhite L., Cisek L., Corden J., Sato M., Maupin P. Actin and myosin biochemistry in relation to cytokinesis. Ann N Y Acad Sci. 1990;582:120–130. doi: 10.1111/j.1749-6632.1990.tb21673.x. [DOI] [PubMed] [Google Scholar]
  45. Rubino S., Fighetti M., Unger E., Cappuccinelli P. Location of actin, myosin, and microtubular structures during directed locomotion of Dictyostelium amebae. J Cell Biol. 1984 Feb;98(2):382–390. doi: 10.1083/jcb.98.2.382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sinard J. H., Pollard T. D. Microinjection into Acanthamoeba castellanii of monoclonal antibodies to myosin-II slows but does not stop cell locomotion. Cell Motil Cytoskeleton. 1989;12(1):42–52. doi: 10.1002/cm.970120106. [DOI] [PubMed] [Google Scholar]
  47. Spudich J. A. In pursuit of myosin function. Cell Regul. 1989 Nov;1(1):1–11. doi: 10.1091/mbc.1.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Swanljung-Collins H., Collins J. H. Ca2+ stimulates the Mg2(+)-ATPase activity of brush border myosin I with three or four calmodulin light chains but inhibits with less than two bound. J Biol Chem. 1991 Jan 15;266(2):1312–1319. [PubMed] [Google Scholar]
  49. Taylor D. L., Blinks J. R., Reynolds G. Contractile basis of ameboid movement. VII. Aequorin luminescence during ameboid movement, endocytosis, and capping. J Cell Biol. 1980 Aug;86(2):599–607. doi: 10.1083/jcb.86.2.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Taylor D. L., Condeelis J. S. Cytoplasmic structure and contractility in amoeboid cells. Int Rev Cytol. 1979;56:57–144. doi: 10.1016/s0074-7696(08)61821-5. [DOI] [PubMed] [Google Scholar]
  51. Taylor D. L., Fechheimer M. Cytoplasmic structure and contractility: the solation--contraction coupling hypothesis. Philos Trans R Soc Lond B Biol Sci. 1982 Nov 4;299(1095):185–197. doi: 10.1098/rstb.1982.0125. [DOI] [PubMed] [Google Scholar]
  52. Tilney L. G., Inoué S. Acrosomal reaction of the Thyone sperm. III. The relationship between actin assembly and water influx during the extension of the acrosomal process. J Cell Biol. 1985 Apr;100(4):1273–1283. doi: 10.1083/jcb.100.4.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Titus M. A., Warrick H. M., Spudich J. A. Multiple actin-based motor genes in Dictyostelium. Cell Regul. 1989 Nov;1(1):55–63. doi: 10.1091/mbc.1.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Tucker R. W., Fay F. S. Distribution of intracellular free calcium in quiescent BALB/c 3T3 cells stimulated by platelet-derived growth factor. Eur J Cell Biol. 1990 Feb;51(1):120–127. [PubMed] [Google Scholar]
  55. Wagner M. C., Barylko B., Albanesi J. P. Tissue distribution and subcellular localization of mammalian myosin I. J Cell Biol. 1992 Oct;119(1):163–170. doi: 10.1083/jcb.119.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wessels D., Soll D. R., Knecht D., Loomis W. F., De Lozanne A., Spudich J. Cell motility and chemotaxis in Dictyostelium amebae lacking myosin heavy chain. Dev Biol. 1988 Jul;128(1):164–177. doi: 10.1016/0012-1606(88)90279-5. [DOI] [PubMed] [Google Scholar]
  57. Yumura S., Mori H., Fukui Y. Localization of actin and myosin for the study of ameboid movement in Dictyostelium using improved immunofluorescence. J Cell Biol. 1984 Sep;99(3):894–899. doi: 10.1083/jcb.99.3.894. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES