Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1989 May 1;108(5):1727–1735. doi: 10.1083/jcb.108.5.1727

Evidence for active interactions between microfilaments and microtubules in myxomycete flagellates

PMCID: PMC2115555  PMID: 2715175

Abstract

We have previously observed the apparent displacement of microfilaments over microtubules in the backbone structure of permeabilized flagellates of Physarum polycephalum upon addition of ATP (Uyeda, T. Q. P., and M. Furuya. 1987. Protoplasma. 140:190-192). We now report that disrupting the microtubular cytoskeleton by treatment with 0.2 mM Ca2+ for 3-30 s inhibits the movement of the microfilaments induced by subsequent treatment with 1 mM Mg-ATP and 10 mM EGTA. Stabilization of microtubules by pretreatment with 50 microM taxol retarded both the disintegrative effect of Ca2+ on the microtubules and the inhibitory effect of Ca2+ on the subsequent, ATP-induced movement of the microfilaments. These results suggest that the movement of the microfilaments depends on the integrity of the microtubular cytoskeleton. EM observation showed that the backbone structure in control permeabilized flagellates consists of two arrays of microtubules closely aligned with bundles of microfilaments of uniform polarity. The microtubular arrays after ATP treatment were no longer associated with microfilaments, yet their alignment was not affected by the ATP treatment. These results imply that the ATP treatment induces reciprocal sliding between the microfilaments and the microtubules, rather than between the microfilaments themselves or between the microtubules themselves. While sliding was best stimulated by ATP, the movement was partially induced by GTP or ATP gamma S, but not by ADP or adenylyl-imidodiphosphate (AMP-PNP). AMP-PNP added in excess to ATP, 50 microM vanadate, or 2 mM erythro-9-[3-(2-hydroxynonyl)]adenine (EHNA) inhibited the sliding. Thus, the pharmacological characteristics of this motility were partly similar to, although not the same as, those of the known microtubule-dependent motilities.

Full Text

The Full Text of this article is available as a PDF (2.7 MB).

Selected References

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

  1. Aldrich H. C. The development of flagella in swarm cells of the myxomycete Physarum flavicomum. J Gen Microbiol. 1968 Feb;50(2):217–222. doi: 10.1099/00221287-50-2-217. [DOI] [PubMed] [Google Scholar]
  2. Begg D. A., Rodewald R., Rebhun L. I. The visualization of actin filament polarity in thin sections. Evidence for the uniform polarity of membrane-associated filaments. J Cell Biol. 1978 Dec;79(3):846–852. doi: 10.1083/jcb.79.3.846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouchard P., Penningroth S. M., Cheung A., Gagnon C., Bardin C. W. erythro-9-[3-(2-Hydroxynonyl)]adenine is an inhibitor of sperm motility that blocks dynein ATPase and protein carboxylmethylase activities. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1033–1036. doi: 10.1073/pnas.78.2.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boyles J., Fox J. E., Phillips D. R., Stenberg P. E. Organization of the cytoskeleton in resting, discoid platelets: preservation of actin filaments by a modified fixation that prevents osmium damage. J Cell Biol. 1985 Oct;101(4):1463–1472. doi: 10.1083/jcb.101.4.1463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Forer A., Jackson W. T., Engberg A. Actin in spindles of Haemanthus katherinae endosperm. II. Distribution of actin in chromosomal spindle fibres, determined by analysis of serial sections. J Cell Sci. 1979 Jun;37:349–371. doi: 10.1242/jcs.37.1.349. [DOI] [PubMed] [Google Scholar]
  6. Gibbons B. H., Gibbons I. R. Flagellar movement and adenosine triphosphatase activity in sea urchin sperm extracted with triton X-100. J Cell Biol. 1972 Jul;54(1):75–97. doi: 10.1083/jcb.54.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gibbons I. R., Cosson M. P., Evans J. A., Gibbons B. H., Houck B., Martinson K. H., Sale W. S., Tang W. J. Potent inhibition of dynein adenosinetriphosphatase and of the motility of cilia and sperm flagella by vanadate. Proc Natl Acad Sci U S A. 1978 May;75(5):2220–2224. doi: 10.1073/pnas.75.5.2220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gorbsky G. J., Sammak P. J., Borisy G. G. Chromosomes move poleward in anaphase along stationary microtubules that coordinately disassemble from their kinetochore ends. J Cell Biol. 1987 Jan;104(1):9–18. doi: 10.1083/jcb.104.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ishikawa H., Bischoff R., Holtzer H. Formation of arrowhead complexes with heavy meromyosin in a variety of cell types. J Cell Biol. 1969 Nov;43(2):312–328. [PMC free article] [PubMed] [Google Scholar]
  10. Kobayashi T., Martensen T., Nath J., Flavin M. Inhibition of dynein ATPase by vanadate, and its possible use as a probe for the role of dynein in cytoplasmic motility. Biochem Biophys Res Commun. 1978 Apr 28;81(4):1313–1318. doi: 10.1016/0006-291x(78)91279-2. [DOI] [PubMed] [Google Scholar]
  11. Koonce M. P., Schliwa M. Reactivation of organelle movements along the cytoskeletal framework of a giant freshwater ameba. J Cell Biol. 1986 Aug;103(2):605–612. doi: 10.1083/jcb.103.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Menzel D., Schliwa M. Motility in the siphonous green alga Bryopsis. I. Spatial organization of the cytoskeleton and organelle movements. Eur J Cell Biol. 1986 Apr;40(2):275–285. [PubMed] [Google Scholar]
  13. Nishida E., Kuwaki T., Sakai H. Phosphorylation of microtubule-associated proteins (MAPs) and pH of the medium control interaction between MAPs and actin filaments. J Biochem. 1981 Aug;90(2):575–578. doi: 10.1093/oxfordjournals.jbchem.a133510. [DOI] [PubMed] [Google Scholar]
  14. Paschal B. M., Vallee R. B. Retrograde transport by the microtubule-associated protein MAP 1C. Nature. 1987 Nov 12;330(6144):181–183. doi: 10.1038/330181a0. [DOI] [PubMed] [Google Scholar]
  15. Penningroth S. M., Cheung A., Olehnik K., Koslosky R. Mechanochemical coupling in the relaxation of rigor-wave sea urchin sperm flagella. J Cell Biol. 1982 Mar;92(3):733–741. doi: 10.1083/jcb.92.3.733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Piperno G. Isolation of a sixth dynein subunit adenosine triphosphatase of Chlamydomonas axonemes. J Cell Biol. 1988 Jan;106(1):133–140. doi: 10.1083/jcb.106.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pollard T. D., Selden S. C., Maupin P. Interaction of actin filaments with microtubules. J Cell Biol. 1984 Jul;99(1 Pt 2):33s–37s. doi: 10.1083/jcb.99.1.33s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Schliwa M., Ezzell R. M., Euteneuer U. erythro-9-[3-(2-Hydroxynonyl)]adenine is an effective inhibitor of cell motility and actin assembly. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6044–6048. doi: 10.1073/pnas.81.19.6044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tiwari S. C., Wick S. M., Williamson R. E., Gunning B. E. Cytoskeleton and integration of cellular function in cells of higher plants. J Cell Biol. 1984 Jul;99(1 Pt 2):63s–69s. doi: 10.1083/jcb.99.1.63s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Uyeda T. Q., Furuya M. Effects of low temperature and calcium on microfilament structure in flagellates of Physarum polycephalum. Exp Cell Res. 1986 Aug;165(2):461–472. doi: 10.1016/0014-4827(86)90599-9. [DOI] [PubMed] [Google Scholar]
  23. Uyeda T. Q., Hatano S., Kohama K., Furuya M. Purification of myxamoebal fragmin, and switching of myxamoebal fragmin to plasmodial fragmin during differentiation of Physarum polycephalum. J Muscle Res Cell Motil. 1988 Jun;9(3):233–240. doi: 10.1007/BF01773893. [DOI] [PubMed] [Google Scholar]
  24. Vale R. D., Reese T. S., Sheetz M. P. Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell. 1985 Aug;42(1):39–50. doi: 10.1016/s0092-8674(85)80099-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vallee R. B., Wall J. S., Paschal B. M., Shpetner H. S. Microtubule-associated protein 1C from brain is a two-headed cytosolic dynein. Nature. 1988 Apr 7;332(6164):561–563. doi: 10.1038/332561a0. [DOI] [PubMed] [Google Scholar]
  26. Weeds A. G., Taylor R. S. Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin. Nature. 1975 Sep 4;257(5521):54–56. doi: 10.1038/257054a0. [DOI] [PubMed] [Google Scholar]
  27. 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