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. 1992 Dec 1;119(5):1193–1203. doi: 10.1083/jcb.119.5.1193

Differential localization of Acanthamoeba myosin I isoforms

PMCID: PMC2289716  PMID: 1447297

Abstract

Acanthamoeba myosins IA and IB were localized by immunofluorescence and immunoelectron microscopy in vegetative and phagocytosing cells and the total cell contents of myosins IA, IB, and IC were quantified by immunoprecipitation. The quantitative distributions of the three myosin I isoforms were then calculated from these data and the previously determined localization of myosin IC. Myosin IA occurs almost exclusively in the cytoplasm, where it accounts for approximately 50% of the total myosin I, in the cortex beneath phagocytic cups and in association with small cytoplasmic vesicles. Myosin IB is the predominant isoform associated with the plasma membrane, large vacuole membranes and phagocytic membranes and accounts for almost half of the total myosin I in the cytoplasm. Myosin IC accounts for a significant fraction of the total myosin I associated with the plasma membrane and large vacuole membranes and is the only myosin I isoform associated with the contractile vacuole membrane. These data suggest that myosin IA may function in cytoplasmic vesicle transport and myosin I-mediated cortical contraction, myosin IB in pseudopod extension and phagocytosis, and myosin IC in contractile vacuole function. In addition, endogenous and exogenously added myosins IA and IB appeared to be associated with the cytoplasmic surface of different subpopulations of purified plasma membranes implying that the different myosin I isoforms are targeted to specific membrane domains through a mechanism that involves more than the affinity of the myosins for anionic phospholipids.

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Selected References

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  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. Baines I. C., Korn E. D. Localization of myosin IC and myosin II in Acanthamoeba castellanii by indirect immunofluorescence and immunogold electron microscopy. J Cell Biol. 1990 Nov;111(5 Pt 1):1895–1904. doi: 10.1083/jcb.111.5.1895. [DOI] [PMC free article] [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. Bowers B., Olszewski T. E., Hyde J. Morphometric analysis of volumes and surface areas in membrane compartments during endocytosis in Acanthamoeba. J Cell Biol. 1981 Mar;88(3):509–515. doi: 10.1083/jcb.88.3.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brzeska H., Lynch T. J., Korn E. D. Localization of the actin-binding sites of Acanthamoeba myosin IB and effect of limited proteolysis on its actin-activated Mg2+-ATPase activity. J Biol Chem. 1988 Jan 5;263(1):427–435. [PubMed] [Google Scholar]
  6. Brzeska H., Lynch T. J., Martin B., Korn E. D. The localization and sequence of the phosphorylation sites of Acanthamoeba myosins I. An improved method for locating the phosphorylated amino acid. J Biol Chem. 1989 Nov 15;264(32):19340–19348. [PubMed] [Google Scholar]
  7. Clarke B. J., Hohman T. C., Bowers B. Purification of plasma membrane from Acanthamoeba castellanii. J Protozool. 1988 Aug;35(3):408–413. doi: 10.1111/j.1550-7408.1988.tb04118.x. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. 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]
  11. Hagen S. J., Kiehart D. P., Kaiser D. A., Pollard T. D. Characterization of monoclonal antibodies to Acanthamoeba myosin-I that cross-react with both myosin-II and low molecular mass nuclear proteins. J Cell Biol. 1986 Dec;103(6 Pt 1):2121–2128. doi: 10.1083/jcb.103.6.2121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hammer J. A., 3rd, Albanesi J. P., Korn E. D. Purification and characterization of a myosin I heavy chain kinase from Acanthamoeba castellanii. J Biol Chem. 1983 Aug 25;258(16):10168–10175. [PubMed] [Google Scholar]
  13. Hammer J. A., 3rd, Korn E. D., Paterson B. M. Acanthamoeba myosin IA, IB, and II heavy chains are synthesized in vitro from Acanthamoeba messenger RNA. J Biol Chem. 1984 Sep 25;259(18):11157–11159. [PubMed] [Google Scholar]
  14. Jung G., Korn E. D., Hammer J. A., 3rd The heavy chain of Acanthamoeba myosin IB is a fusion of myosin-like and non-myosin-like sequences. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6720–6724. doi: 10.1073/pnas.84.19.6720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kessler S. W. Use of protein A-bearing staphylococci for the immunoprecipitation and isolation of antigens from cells. Methods Enzymol. 1981;73(Pt B):442–459. doi: 10.1016/0076-6879(81)73084-2. [DOI] [PubMed] [Google Scholar]
  16. Korn E. D., Atkinson M. A., Brzeska H., Hammer J. A., 3rd, Jung G., Lynch T. J. Structure-function studies on Acanthamoeba myosins IA, IB, and II. J Cell Biochem. 1988 Jan;36(1):37–50. doi: 10.1002/jcb.240360105. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Korn E. D., Hammer J. A., 3rd Myosins of nonmuscle cells. Annu Rev Biophys Biophys Chem. 1988;17:23–45. doi: 10.1146/annurev.bb.17.060188.000323. [DOI] [PubMed] [Google Scholar]
  19. Kulesza-Lipka D., Baines I. C., Brzeska H., Korn E. D. Immunolocalization of myosin I heavy chain kinase in Acanthamoeba castellanii and binding of purified kinase to isolated plasma membranes. J Cell Biol. 1991 Oct;115(1):109–119. doi: 10.1083/jcb.115.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Larson R. E., Espindola F. S., Espreafico E. M. Calmodulin-binding proteins and calcium/calmodulin-regulated enzyme activities associated with brain actomyosin. J Neurochem. 1990 Apr;54(4):1288–1294. doi: 10.1111/j.1471-4159.1990.tb01961.x. [DOI] [PubMed] [Google Scholar]
  22. Larson R. E., Pitta D. E., Ferro J. A. A novel 190 kDa calmodulin-binding protein associated with brain actomyosin. Braz J Med Biol Res. 1988;21(2):213–217. [PubMed] [Google Scholar]
  23. Lynch T. J., Albanesi J. P., Korn E. D., Robinson E. A., Bowers B., Fujisaki H. ATPase activities and actin-binding properties of subfragments of Acanthamoeba myosin IA. J Biol Chem. 1986 Dec 25;261(36):17156–17162. [PubMed] [Google Scholar]
  24. Lynch T. J., Brzeska H., Baines I. C., Korn E. D. Purification of myosin I and myosin I heavy chain kinase from Acanthamoeba castellanii. Methods Enzymol. 1991;196:12–23. doi: 10.1016/0076-6879(91)96004-b. [DOI] [PubMed] [Google Scholar]
  25. Lynch T. J., Brzeska H., Miyata H., Korn E. D. Purification and characterization of a third isoform of myosin I from Acanthamoeba castellanii. J Biol Chem. 1989 Nov 15;264(32):19333–19339. [PubMed] [Google Scholar]
  26. Mercer J. A., Seperack P. K., Strobel M. C., Copeland N. G., Jenkins N. A. Novel myosin heavy chain encoded by murine dilute coat colour locus. Nature. 1991 Feb 21;349(6311):709–713. doi: 10.1038/349709a0. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Montell C., Rubin G. M. The Drosophila ninaC locus encodes two photoreceptor cell specific proteins with domains homologous to protein kinases and the myosin heavy chain head. Cell. 1988 Mar 11;52(5):757–772. doi: 10.1016/0092-8674(88)90413-8. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Pollard T. D., Korn E. D. Acanthamoeba myosin. I. Isolation from Acanthamoeba castellanii of an enzyme similar to muscle myosin. J Biol Chem. 1973 Jul 10;248(13):4682–4690. [PubMed] [Google Scholar]
  31. Prendergast J. A., Murray L. E., Rowley A., Carruthers D. R., Singer R. A., Johnston G. C. Size selection identifies new genes that regulate Saccharomyces cerevisiae cell proliferation. Genetics. 1990 Jan;124(1):81–90. doi: 10.1093/genetics/124.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]

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