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. 1993 Jan 1;120(1):163–173. doi: 10.1083/jcb.120.1.163

Dictyostelium mutants lacking the cytoskeletal protein coronin are defective in cytokinesis and cell motility

PMCID: PMC2119478  PMID: 8380174

Abstract

Coronin is an actin-binding protein in Dictyostelium discoideum that is enriched at the leading edge of the cells and in projections of the cell surface called crowns. The polypeptide sequence of coronin is distinguished by its similarities to the beta-subunits of trimeric G proteins (E. L. de Hostos, B. Bradtke, F. Lottspeich, R. Guggenheim, and G. Gerisch, 1991. EMBO (Eur. Mol. Biol. Organ.) J. 10:4097-4104). To elucidate the in vivo function of coronin, null mutants have been generated by gene replacement. The mutant cells lacking coronin grow and migrate more slowly than wild-type cells. When these cor- cells grow in liquid medium they become multinucleate, indicating a role of coronin in cytokinesis. To explore this role, coronin has been localized in mitotic wild-type cells by immunofluorescence labeling. During separation of the daughter cells, coronin is strongly accumulated at their distal portions including the leading edges. This contrasts with the localization of myosin II in the cleavage furrow and suggests that coronin functions independently of the conventional myosin in facilitating cytokinesis.

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Selected References

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  1. André E., Brink M., Gerisch G., Isenberg G., Noegel A., Schleicher M., Segall J. E., Wallraff E. A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis. J Cell Biol. 1989 Mar;108(3):985–995. doi: 10.1083/jcb.108.3.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashworth J. M., Watts D. J. Metabolism of the cellular slime mould Dictyostelium discoideum grown in axenic culture. Biochem J. 1970 Sep;119(2):175–182. doi: 10.1042/bj1190175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barr F. A., Leyte A., Huttner W. B. Trimeric G proteins and vesicle formation. Trends Cell Biol. 1992 Apr;2(4):91–94. doi: 10.1016/0962-8924(92)90001-4. [DOI] [PubMed] [Google Scholar]
  4. Berlot C. H., Spudich J. A., Devreotes P. N. Chemoattractant-elicited increases in myosin phosphorylation in Dictyostelium. Cell. 1985 Nov;43(1):307–314. doi: 10.1016/0092-8674(85)90036-4. [DOI] [PubMed] [Google Scholar]
  5. Bomblies L., Biegelmann E., Döring V., Gerisch G., Krafft-Czepa H., Noegel A. A., Schleicher M., Humbel B. M. Membrane-enclosed crystals in Dictyostelium discoideum cells, consisting of developmentally regulated proteins with sequence similarities to known esterases. J Cell Biol. 1990 Mar;110(3):669–679. doi: 10.1083/jcb.110.3.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brink M., Gerisch G., Isenberg G., Noegel A. A., Segall J. E., Wallraff E., Schleicher M. A Dictyostelium mutant lacking an F-actin cross-linking protein, the 120-kD gelation factor. J Cell Biol. 1990 Oct;111(4):1477–1489. doi: 10.1083/jcb.111.4.1477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burgoyne R. D. Trimeric G proteins in Golgi transport. Trends Biochem Sci. 1992 Mar;17(3):87–88. [PubMed] [Google Scholar]
  8. Choi W. J., Clark M. W., Chen J. X., Jong A. Y. The CDC4 gene product is associated with the yeast nuclear skeleton. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1324–1330. doi: 10.1016/0006-291x(90)91594-i. [DOI] [PubMed] [Google Scholar]
  9. Claviez M., Pagh K., Maruta H., Baltes W., Fisher P., Gerisch G. Electron microscopic mapping of monoclonal antibodies on the tail region of Dictyostelium myosin. EMBO J. 1982;1(8):1017–1022. doi: 10.1002/j.1460-2075.1982.tb01287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cox D., Condeelis J., Wessels D., Soll D., Kern H., Knecht D. A. Targeted disruption of the ABP-120 gene leads to cells with altered motility. J Cell Biol. 1992 Feb;116(4):943–955. doi: 10.1083/jcb.116.4.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cunningham C. C., Gorlin J. B., Kwiatkowski D. J., Hartwig J. H., Janmey P. A., Byers H. R., Stossel T. P. Actin-binding protein requirement for cortical stability and efficient locomotion. Science. 1992 Jan 17;255(5042):325–327. doi: 10.1126/science.1549777. [DOI] [PubMed] [Google Scholar]
  12. Cunningham C. C., Stossel T. P., Kwiatkowski D. J. Enhanced motility in NIH 3T3 fibroblasts that overexpress gelsolin. Science. 1991 Mar 8;251(4998):1233–1236. doi: 10.1126/science.1848726. [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. Devreotes P. N., Zigmond S. H. Chemotaxis in eukaryotic cells: a focus on leukocytes and Dictyostelium. Annu Rev Cell Biol. 1988;4:649–686. doi: 10.1146/annurev.cb.04.110188.003245. [DOI] [PubMed] [Google Scholar]
  15. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  16. Fisher P. R., Grant W. N., Dohrmann U., Williams K. L. Spontaneous turning behaviour by Dictyostelium discoideum slugs. J Cell Sci. 1983 Jul;62:161–170. doi: 10.1242/jcs.62.1.161. [DOI] [PubMed] [Google Scholar]
  17. Fisher P. R., Merkl R., Gerisch G. Quantitative analysis of cell motility and chemotaxis in Dictyostelium discoideum by using an image processing system and a novel chemotaxis chamber providing stationary chemical gradients. J Cell Biol. 1989 Mar;108(3):973–984. doi: 10.1083/jcb.108.3.973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fisher P. R., Smith E., Williams K. L. An extracellular chemical signal controlling phototactic behavior by D. discoideum slugs. Cell. 1981 Mar;23(3):799–807. doi: 10.1016/0092-8674(81)90444-x. [DOI] [PubMed] [Google Scholar]
  19. Fukui Y., De Lozanne A., Spudich J. A. Structure and function of the cytoskeleton of a Dictyostelium myosin-defective mutant. J Cell Biol. 1990 Feb;110(2):367–378. doi: 10.1083/jcb.110.2.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Fukui Y., Yumura S., Yumura T. K. Agar-overlay immunofluorescence: high-resolution studies of cytoskeletal components and their changes during chemotaxis. Methods Cell Biol. 1987;28:347–356. doi: 10.1016/s0091-679x(08)61655-6. [DOI] [PubMed] [Google Scholar]
  22. Gerisch G., Noegel A. A., Schleicher M. Genetic alteration of proteins in actin-based motility systems. Annu Rev Physiol. 1991;53:607–628. doi: 10.1146/annurev.ph.53.030191.003135. [DOI] [PubMed] [Google Scholar]
  23. Jay P. Y., Elson E. L. Surface particle transport mechanism independent of myosin II in Dictyostelium. Nature. 1992 Apr 2;356(6368):438–440. doi: 10.1038/356438a0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Knecht D. A., Cohen S. M., Loomis W. F., Lodish H. F. Developmental regulation of Dictyostelium discoideum actin gene fusions carried on low-copy and high-copy transformation vectors. Mol Cell Biol. 1986 Nov;6(11):3973–3983. doi: 10.1128/mcb.6.11.3973-3983.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Knecht D. A., Loomis W. F. Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum. Science. 1987 May 29;236(4805):1081–1086. doi: 10.1126/science.3576221. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Mehdy M. C., Ratner D., Firtel R. A. Induction and modulation of cell-type-specific gene expression in Dictyostelium. Cell. 1983 Mar;32(3):763–771. doi: 10.1016/0092-8674(83)90062-4. [DOI] [PubMed] [Google Scholar]
  30. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  31. Satterwhite L. L., Pollard T. D. Cytokinesis. Curr Opin Cell Biol. 1992 Feb;4(1):43–52. doi: 10.1016/0955-0674(92)90057-j. [DOI] [PubMed] [Google Scholar]
  32. Schleicher M., Noegel A. A. Dynamics of the Dictyostelium cytoskeleton during chemotaxis. New Biol. 1992 May;4(5):461–472. [PubMed] [Google Scholar]
  33. Segall J. E., Fisher P. R., Gerisch G. Selection of chemotaxis mutants of Dictyostelium discoideum. J Cell Biol. 1987 Jan;104(1):151–161. doi: 10.1083/jcb.104.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Waddell D. R., Duffy K., Vogel G. Cytokinesis is defective in Dictyostelium mutants with altered phagocytic recognition, adhesion, and vegetative cell cohesion properties. J Cell Biol. 1987 Nov;105(5):2293–2300. doi: 10.1083/jcb.105.5.2293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wallraff E., Schleicher M., Modersitzki M., Rieger D., Isenberg G., Gerisch G. Selection of Dictyostelium mutants defective in cytoskeletal proteins: use of an antibody that binds to the ends of alpha-actinin rods. EMBO J. 1986 Jan;5(1):61–67. doi: 10.1002/j.1460-2075.1986.tb04178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Wessels D., Soll D. R. Myosin II heavy chain null mutant of Dictyostelium exhibits defective intracellular particle movement. J Cell Biol. 1990 Sep;111(3):1137–1148. doi: 10.1083/jcb.111.3.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Witke W., Nellen W., Noegel A. Homologous recombination in the Dictyostelium alpha-actinin gene leads to an altered mRNA and lack of the protein. EMBO J. 1987 Dec 20;6(13):4143–4148. doi: 10.1002/j.1460-2075.1987.tb02760.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Witke W., Schleicher M., Noegel A. A. Redundancy in the microfilament system: abnormal development of Dictyostelium cells lacking two F-actin cross-linking proteins. Cell. 1992 Jan 10;68(1):53–62. doi: 10.1016/0092-8674(92)90205-q. [DOI] [PubMed] [Google Scholar]
  41. Yochem J., Byers B. Structural comparison of the yeast cell division cycle gene CDC4 and a related pseudogene. J Mol Biol. 1987 May 20;195(2):233–245. doi: 10.1016/0022-2836(87)90646-2. [DOI] [PubMed] [Google Scholar]
  42. de Hostos E. L., Bradtke B., Lottspeich F., Guggenheim R., Gerisch G. Coronin, an actin binding protein of Dictyostelium discoideum localized to cell surface projections, has sequence similarities to G protein beta subunits. EMBO J. 1991 Dec;10(13):4097–4104. doi: 10.1002/j.1460-2075.1991.tb04986.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. van der Voorn L., Gebbink M., Plasterk R. H., Ploegh H. L. Characterization of a G-protein beta-subunit gene from the nematode Caenorhabditis elegans. J Mol Biol. 1990 May 5;213(1):17–26. doi: 10.1016/s0022-2836(05)80118-4. [DOI] [PubMed] [Google Scholar]

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