Skip to main content
NCBI home page
Cell Regulation logoLink to Cell Regulation
. 1990 Sep;1(10):715–729. doi: 10.1091/mbc.1.10.715

Regulation of slug size by the cell adhesion molecule gp80 in Dictyostelium discoideum.

R K Kamboj 1, T Y Lam 1, C H Siu 1
PMCID: PMC361651  PMID: 1966011

Abstract

We previously provided in vitro evidence that the cell surface glycoprotein of Mr80,000 (gp80) of Dictyostelium discoideum is capable of mediating EDTA-resistant cell-cell binding. Expression of gp80 is specific for the aggregation stage when cells form tight aggregates. To investigate the physiological role of gp80, Dictyostelium cells were transformed with a vector containing gp80 cDNA fused to an actin promoter. gp80 transcripts were detected in transformed cells in their vegetative growth phase. Transformants at this stage also exhibited EDTA-resistant cell cohesion, thus providing direct in vivo evidence that gp80 mediates cell-cell binding via homophilic interaction. While aggregates of the parental strain KAX3 had the tendency to break up to form small slugs, transformants expressing an increased amount of gp80 were able to maintain the integrity of aggregates, giving rise to larger slugs, resulting in the formation of bigger fruiting bodies. To further demonstrate that the increase in slug size could be correlated with the expression of gp80, cells of the parental strain were treated with exogenous cAMP pulses to stimulate an over-expression of gp80. The treated cells also gave rise to larger slugs, consistent with the notion that slug size is influenced by intercellular adhesiveness during development.

Full text

PDF
715

Images in this article

717Image
on p.717
718Image
on p.718
720Image
on p.720
721Image
on p.721
723Image
on p.723

Selected References

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

  1. Beug H., Katz F. E., Gerisch G. Dynamics of antigenic membrane sites relating to cell aggregation in Dictyostelium discoideum. J Cell Biol. 1973 Mar;56(3):647–658. doi: 10.1083/jcb.56.3.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chisholm R. L., Barklis E., Lodish H. F. Mechanism of sequential induction of cell-type specific mRNAs in Dictyostelium differentiation. Nature. 1984 Jul 5;310(5972):67–69. doi: 10.1038/310067a0. [DOI] [PubMed] [Google Scholar]
  3. Choi A. H., Siu C. H. Filopodia are enriched in a cell cohesion molecule of Mr 80,000 and participate in cell-cell contact formation in Dictyostelium discoideum. J Cell Biol. 1987 May;104(5):1375–1387. doi: 10.1083/jcb.104.5.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chung S., Landfear S. M., Blumberg D. D., Cohen N. S., Lodish H. F. Synthesis and stability of developmentally regulated dictyostelium mRNAs are affected by cell--cell contact and cAMP. Cell. 1981 Jun;24(3):785–797. doi: 10.1016/0092-8674(81)90104-5. [DOI] [PubMed] [Google Scholar]
  5. Cocucci S. M., Sussman M. RNA in cytoplasmic and nuclear fractions of cellular slime mold amebas. J Cell Biol. 1970 May;45(2):399–407. doi: 10.1083/jcb.45.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cunningham B. A., Hemperly J. J., Murray B. A., Prediger E. A., Brackenbury R., Edelman G. M. Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing. Science. 1987 May 15;236(4803):799–806. doi: 10.1126/science.3576199. [DOI] [PubMed] [Google Scholar]
  7. De Silva N. S., Siu C. H. Preferential incorporation of phospholipids into plasma membranes during cell aggregation of Dictyostelium discoideum. J Biol Chem. 1980 Sep 25;255(18):8489–8496. [PubMed] [Google Scholar]
  8. Edelman G. M. Cell adhesion molecules in the regulation of animal form and tissue pattern. Annu Rev Cell Biol. 1986;2:81–116. doi: 10.1146/annurev.cb.02.110186.000501. [DOI] [PubMed] [Google Scholar]
  9. Edelman G. M., Murray B. A., Mege R. M., Cunningham B. A., Gallin W. J. Cellular expression of liver and neural cell adhesion molecules after transfection with their cDNAs results in specific cell-cell binding. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8502–8506. doi: 10.1073/pnas.84.23.8502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Faure M., Podgorski G. J., Franke J., Kessin R. H. Disruption of Dictyostelium discoideum morphogenesis by overproduction of cAMP phosphodiesterase. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8076–8080. doi: 10.1073/pnas.85.21.8076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feinberg A. P., Springer W. R., Barondes S. H. Segregation of pre-stalk and pre-spore cells of Dictyostelium discoideum: observations consistent with selective cell cohesion. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3977–3981. doi: 10.1073/pnas.76.8.3977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  13. Garrod D. R. Acquisition of cohesiveness by slime mould cells prior to morphogenesis. Exp Cell Res. 1972 Jun;72(2):588–591. doi: 10.1016/0014-4827(72)90041-9. [DOI] [PubMed] [Google Scholar]
  14. Geltosky J. E., Weseman J., Bakke A., Lerner R. A. Identification of a cell surface glycoprotein involved in cell aggregation in D. discoideum. Cell. 1979 Oct;18(2):391–398. doi: 10.1016/0092-8674(79)90058-8. [DOI] [PubMed] [Google Scholar]
  15. Gerisch G. Univalent antibody fragments as tools for the analysis of cell interactions in Dictyostelium. Curr Top Dev Biol. 1980;14(Pt 2):243–270. doi: 10.1016/s0070-2153(08)60197-0. [DOI] [PubMed] [Google Scholar]
  16. Gerisch G., Weinhart U., Bertholdt G., Claviez M., Stadler J. Incomplete contact site A glycoprotein in HL220, a modB mutant of Dictyostelium discoideum. J Cell Sci. 1985 Feb;73:49–68. doi: 10.1242/jcs.73.1.49. [DOI] [PubMed] [Google Scholar]
  17. Harloff C., Gerisch G., Noegel A. A. Selective elimination of the contact site A protein of Dictyostelium discoideum by gene disruption. Genes Dev. 1989 Dec;3(12A):2011–2019. doi: 10.1101/gad.3.12a.2011. [DOI] [PubMed] [Google Scholar]
  18. Hohmann H. P., Bozzaro S., Merkl R., Wallraff E., Yoshida M., Weinhart U., Gerisch G. Post-translational glycosylation of the contact site A protein of Dictyostelium discoideum is important for stability but not for its function in cell adhesion. EMBO J. 1987 Dec 1;6(12):3663–3671. doi: 10.1002/j.1460-2075.1987.tb02699.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kamboj R. K., Gariepy J., Siu C. H. Identification of an octapeptide involved in homophilic interaction of the cell adhesion molecule gp80 of dictyostelium discoideum. Cell. 1989 Nov 17;59(4):615–625. doi: 10.1016/0092-8674(89)90007-x. [DOI] [PubMed] [Google Scholar]
  20. Kamboj R. K., Siu C. H. Mapping of the monoclonal antibody 80L5C4 epitope on the cell adhesion molecule gp80 of Dictyostelium discoideum. Biochim Biophys Acta. 1988 Nov 10;951(1):78–84. doi: 10.1016/0167-4781(88)90027-9. [DOI] [PubMed] [Google Scholar]
  21. Kamboj R. K., Wong L. M., Lam T. Y., Siu C. H. Mapping of a cell-binding domain in the cell adhesion molecule gp80 of Dictyostelium discoideum. J Cell Biol. 1988 Nov;107(5):1835–1843. doi: 10.1083/jcb.107.5.1835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kopachik W. Size regulation in Dictyostelium. J Embryol Exp Morphol. 1982 Apr;68:23–35. [PubMed] [Google Scholar]
  24. Kumagai A., Pupillo M., Gundersen R., Miake-Lye R., Devreotes P. N., Firtel R. A. Regulation and function of G alpha protein subunits in Dictyostelium. Cell. 1989 Apr 21;57(2):265–275. doi: 10.1016/0092-8674(89)90964-1. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Lam T. Y., Siu C. H. Synthesis of stage-specific glycoproteins in Dictyostelium discoideum during development. Dev Biol. 1981 Apr 15;83(1):127–137. doi: 10.1016/s0012-1606(81)80015-2. [DOI] [PubMed] [Google Scholar]
  27. Loomis W. F. Cell-cell adhesion in Dictyostelium discoideum. Dev Genet. 1988;9(4-5):549–559. doi: 10.1002/dvg.1020090431. [DOI] [PubMed] [Google Scholar]
  28. Ma P. C., Siu C. H. A pharmacologically distinct cyclic AMP receptor is responsible for the regulation of gp80 expression in Dictyostelium discoideum. Mol Cell Biol. 1990 Jul;10(7):3297–3306. doi: 10.1128/mcb.10.7.3297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mann S. K., Firtel R. A. Two-phase regulatory pathway controls cAMP receptor-mediated expression of early genes in Dictyostelium. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1924–1928. doi: 10.1073/pnas.86.6.1924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. McClay D. R., Ettensohn C. A. Cell adhesion in morphogenesis. Annu Rev Cell Biol. 1987;3:319–345. doi: 10.1146/annurev.cb.03.110187.001535. [DOI] [PubMed] [Google Scholar]
  31. Murray B. A., Wheeler S., Jongens T., Loomis W. F. Mutations affecting a surface glycoprotein, gp80, of Dictyostelium discoideum. Mol Cell Biol. 1984 Mar;4(3):514–519. doi: 10.1128/mcb.4.3.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Murray B. A., Yee L. D., Loomis W. F. Immunological analysis of glycoprotein (contact sites A) involved in intercellular adhesion of Dictyostelium discoideum. J Supramol Struct Cell Biochem. 1981;17(3):197–211. doi: 10.1002/jsscb.380170302. [DOI] [PubMed] [Google Scholar]
  33. Müller K., Gerisch G. A specific glycoprotein as the target site of adhesion blocking Fab in aggregating Dictyostelium cells. Nature. 1978 Aug 3;274(5670):445–449. doi: 10.1038/274445a0. [DOI] [PubMed] [Google Scholar]
  34. Nagafuchi A., Shirayoshi Y., Okazaki K., Yasuda K., Takeichi M. Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA. Nature. 1987 Sep 24;329(6137):341–343. doi: 10.1038/329341a0. [DOI] [PubMed] [Google Scholar]
  35. Nellen W., Silan C., Firtel R. A. DNA-mediated transformation in Dictyostelium discoideum: regulated expression of an actin gene fusion. Mol Cell Biol. 1984 Dec;4(12):2890–2898. doi: 10.1128/mcb.4.12.2890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nellen W., Silan C., Saur U., Firtel R. A. Regulatory sequences in the promoter of the Dictyostelium Actin 6 gene. EMBO J. 1986 Dec 1;5(12):3367–3372. doi: 10.1002/j.1460-2075.1986.tb04652.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Noegel A., Gerisch G., Stadler J., Westphal M. Complete sequence and transcript regulation of a cell adhesion protein from aggregating Dictyostelium cells. EMBO J. 1986 Jul;5(7):1473–1476. doi: 10.1002/j.1460-2075.1986.tb04384.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Parish R. W., Schmidlin S. Synthesis of plasma membrane proteins during development of Dictyostelium discoideum. FEBS Lett. 1979 Feb 15;98(2):251–256. doi: 10.1016/0014-5793(79)80193-3. [DOI] [PubMed] [Google Scholar]
  39. Poole S. J., Firtel R. A. Genomic instability and mobile genetic elements in regions surrounding two discoidin I genes of Dictyostelium discoideum. Mol Cell Biol. 1984 Apr;4(4):671–680. doi: 10.1128/mcb.4.4.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Romans P., Firtel R. A., Saxe C. L., 3rd Gene-specific expression of the actin multigene family of Dictyostelium discoideum. J Mol Biol. 1985 Nov 20;186(2):337–355. doi: 10.1016/0022-2836(85)90109-3. [DOI] [PubMed] [Google Scholar]
  41. Simon M. N., Driscoll D., Mutzel R., Part D., Williams J., Véron M. Overproduction of the regulatory subunit of the cAMP-dependent protein kinase blocks the differentiation of Dictyostelium discoideum. EMBO J. 1989 Jul;8(7):2039–2043. doi: 10.1002/j.1460-2075.1989.tb03612.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Siu C. H., Cho A., Choi A. H. The contact site A glycoprotein mediates cell-cell adhesion by homophilic binding in Dictyostelium discoideum. J Cell Biol. 1987 Dec;105(6 Pt 1):2523–2533. doi: 10.1083/jcb.105.6.2523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Siu C. H., Des Roches B., Lam T. Y. Involvement of a cell-surface glycoprotein in the cell-sorting process of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6596–6600. doi: 10.1073/pnas.80.21.6596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Siu C. H., Lam T. Y., Choi A. H. Inhibition of cell-cell binding at the aggregation stage of Dictyostelium discoideum development by monoclonal antibodies directed against an 80,000-dalton surface glycoprotein. J Biol Chem. 1985 Dec 15;260(29):16030–16036. [PubMed] [Google Scholar]
  45. Siu C. H., Lam T. Y. Mediation of cell-cell adhesion by the altered contact site A glycoprotein expressed in modB mutants of Dictyostelium discoideum. Exp Cell Res. 1988 Aug;177(2):338–346. doi: 10.1016/0014-4827(88)90467-3. [DOI] [PubMed] [Google Scholar]
  46. Siu C. H., Lam T. Y., Wong L. M. Expression of the contact site A glycoprotein in Dictyostelium discoideum: quantitation and developmental regulation. Biochim Biophys Acta. 1988 Mar 11;968(3):283–290. doi: 10.1016/0167-4889(88)90019-5. [DOI] [PubMed] [Google Scholar]
  47. Siu C. H., Wong L. M., Lam T. Y., Kamboj R. K., Choi A., Cho A. Molecular mechanisms of cell-cell interaction in Dictyostelium discoideum. Biochem Cell Biol. 1988 Oct;66(10):1089–1099. doi: 10.1139/o88-126. [DOI] [PubMed] [Google Scholar]
  48. Stadler J., Keenan T. W., Bauer G., Gerisch G. The contact site A glycoprotein of Dictyostelium discoideum carries a phospholipid anchor of a novel type. EMBO J. 1989 Feb;8(2):371–377. doi: 10.1002/j.1460-2075.1989.tb03387.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tasaka M., Takeuchi I. Sorting out behaviour of disaggregated cells in the absence of morphogenesis in Dictyostelium discoideum. J Embryol Exp Morphol. 1979 Jan;49:89–102. [PubMed] [Google Scholar]
  50. 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]
  51. Williams A. F., Barclay A. N. The immunoglobulin superfamily--domains for cell surface recognition. Annu Rev Immunol. 1988;6:381–405. doi: 10.1146/annurev.iy.06.040188.002121. [DOI] [PubMed] [Google Scholar]
  52. 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]
  53. Wong L. M., Siu C. H. Cloning of cDNA for the contact site A glycoprotein of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4248–4252. doi: 10.1073/pnas.83.12.4248. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Cell Regulation are provided here courtesy of American Society for Cell Biology

RESOURCES