Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Nov 1;103(5):1999–2015. doi: 10.1083/jcb.103.5.1999

Cellular and subcellular distribution of a cAMP-regulated prestalk protein and prespore protein in Dictyostelium discoideum: a study on the ontogeny of prestalk and prespore cells

PMCID: PMC2114392  PMID: 3023394

Abstract

We have analyzed a developmentally and spatially regulated prestalk- specific gene and a prespore-specific gene from Dictyostelium. The prestalk gene, pst-cathepsin, encodes a protein highly homologous to the lysosomal cysteine proteinases cathepsin H and cathepsin B. The prespore gene encodes a protein with some homology to the anti- bacterial toxin crambin and has been designated beejin. Using the lambda gtll system, we have made polyclonal antibodies directed against a portion of the protein encoded by pst-cathepsin and other antibodies directed against the beejin protein. Both antibodies stain single bands on Western blots. By immunofluorescence and Western blots, pst- cathepsin is not present in vegetative cells or developing cells during the first approximately 10 h of development. It then appears with a punctate distribution in a subset of developing cells. Beejin is detected only after approximately 15 h of development, also in a subset of cells. Pst-cathepsin is distributed in the anterior approximately 1/10 of migrating slugs and on the peripheral posterior surfaces of slugs. Beejin is distributed in the posterior region of slugs. Expression of both pst-cathepsin and beejin can be induced in subsets of isolated cultured cells by a combination of conditioned medium and extracellular cAMP in agreement with the regulation of the mRNAs encoding these proteins. We have used the antibodies as markers for cell type to examine the ontogeny and the spatial distribution of prestalk and prespore cells throughout multicellular development. Our findings suggest that prestalk cell differentiation is independent of position within the aggregate and that the spatial localization of prestalk cells within the multicellular aggregate arises from sorting of the prestalk cells after their induction. We have also found a class of cell in developing aggregates that contains neither the prestalk nor the prespore markers.

Full Text

The Full Text of this article is available as a PDF (6.9 MB).

Selected References

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

  1. Armant D. R., Rutherford C. L. 5'-AMP nucleotidase is localized in the area of cell-cell contact of prespore and prestalk regions during culmination of Dictyostelium discoideum. Mech Ageing Dev. 1979 May;10(3-4):199–217. doi: 10.1016/0047-6374(79)90034-4. [DOI] [PubMed] [Google Scholar]
  2. Barklis E., Lodish H. F. Regulation of dictyostelium discoideum mRNAs specific for prespore or prestalk cells. Cell. 1983 Apr;32(4):1139–1148. doi: 10.1016/0092-8674(83)90297-0. [DOI] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blose S. H., Matsumura F., Lin J. J. Structure of vimentin 10-nm filaments probed with a monoclonal antibody that recognizes a common antigenic determinant on vimentin and tropomyosin. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):455–463. doi: 10.1101/sqb.1982.046.01.042. [DOI] [PubMed] [Google Scholar]
  5. Bonner J. T. EVIDENCE FOR THE SORTING OUT OF CELLS IN THE DEVELOPMENT OF THE CELLULAR SLIME MOLDS. Proc Natl Acad Sci U S A. 1959 Mar;45(3):379–384. doi: 10.1073/pnas.45.3.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Danto S. I., Fischman D. A. Immunocytochemical analysis of intermediate filaments in embryonic heart cells with monoclonal antibodies to desmin. J Cell Biol. 1984 Jun;98(6):2179–2191. doi: 10.1083/jcb.98.6.2179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Datta S., Gomer R. H., Firtel R. A. Spatial and temporal regulation of a foreign gene by a prestalk-specific promoter in transformed Dictyostelium discoideum. Mol Cell Biol. 1986 Mar;6(3):811–820. doi: 10.1128/mcb.6.3.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Devine K. M., Bergmann J. E., Loomis W. F. Spore coat proteins of Dictyostelium discoideum are packaged in prespore vesicles. Dev Biol. 1983 Oct;99(2):437–446. doi: 10.1016/0012-1606(83)90293-2. [DOI] [PubMed] [Google Scholar]
  9. Devine K. M., Loomis W. F. Molecular characterization of anterior-like cells in Dictyostelium discoideum. Dev Biol. 1985 Feb;107(2):364–372. doi: 10.1016/0012-1606(85)90318-5. [DOI] [PubMed] [Google Scholar]
  10. Doolittle R. F. Similar amino acid sequences: chance or common ancestry? Science. 1981 Oct 9;214(4517):149–159. doi: 10.1126/science.7280687. [DOI] [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. Firtel R. A., Bonner J. Developmental control of -1-4 glucan phosphorylase in the cellular slime mold Dictyostelium discoideum. Dev Biol. 1972 Sep;29(1):85–103. doi: 10.1016/0012-1606(72)90046-2. [DOI] [PubMed] [Google Scholar]
  13. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  14. Franke W. W., Schmid E., Wellsteed J., Grund C., Gigi O., Geiger B. Change of cytokeratin filament organization during the cell cycle: selective masking of an immunologic determinant in interphase PtK2 cells. J Cell Biol. 1983 Oct;97(4):1255–1260. doi: 10.1083/jcb.97.4.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gomer R. H., Datta S., Mehdy M., Crowley T., Sivertsen A., Nellen W., Reymond C., Mann S., Firtel R. A. Regulation of cell-type-specific gene expression in Dictyostelium. Cold Spring Harb Symp Quant Biol. 1985;50:801–812. doi: 10.1101/sqb.1985.050.01.098. [DOI] [PubMed] [Google Scholar]
  16. Gomer R. H., Lazarides E. The synthesis and deployment of filamin in chicken skeletal muscle. Cell. 1981 Feb;23(2):524–532. doi: 10.1016/0092-8674(81)90148-3. [DOI] [PubMed] [Google Scholar]
  17. Grant W. N., Williams K. L. Monoclonal antibody characterisation of slime sheath: the extracellular matrix of Dictyostelium discoideum. EMBO J. 1983;2(6):935–940. doi: 10.1002/j.1460-2075.1983.tb01524.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hunkapiller M. W., Lujan E., Ostrander F., Hood L. E. Isolation of microgram quantities of proteins from polyacrylamide gels for amino acid sequence analysis. Methods Enzymol. 1983;91:227–236. doi: 10.1016/s0076-6879(83)91019-4. [DOI] [PubMed] [Google Scholar]
  19. Johnson G. D., Nogueira Araujo G. M. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981;43(3):349–350. doi: 10.1016/0022-1759(81)90183-6. [DOI] [PubMed] [Google Scholar]
  20. Kimmel A. R., Firtel R. A. Sequence organization in Dictyostelium: unique structure at the 5'-ends of protein coding genes. Nucleic Acids Res. 1983 Jan 25;11(2):541–552. doi: 10.1093/nar/11.2.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Knecht D. A., Dimond R. L., Wheeler S., Loomis W. F. Antigenic determinants shared by lysosomal proteins of Dictyostelium discoideum. Characterization using monoclonal antibodies and isolation of mutations affecting the determinant. J Biol Chem. 1984 Aug 25;259(16):10633–10640. [PubMed] [Google Scholar]
  22. Loomis W. F., Murray B. A., Yee L., Jongens T. Adhesion-blocking antibodies prepared against gp150 react with gp80 of Dictyostelium. Exp Cell Res. 1983 Aug;147(1):231–234. doi: 10.1016/0014-4827(83)90290-2. [DOI] [PubMed] [Google Scholar]
  23. MacWilliams H. K., Bonner J. T. The prestalk-prespore pattern in cellular slime molds. Differentiation. 1979;14(1-2):1–22. doi: 10.1111/j.1432-0436.1979.tb01006.x. [DOI] [PubMed] [Google Scholar]
  24. Max E. E., Battey J., Ney R., Kirsch I. R., Leder P. Duplication and deletion in the human immunoglobulin epsilon genes. Cell. 1982 Jun;29(2):691–699. doi: 10.1016/0092-8674(82)90185-4. [DOI] [PubMed] [Google Scholar]
  25. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  26. Mehdy M. C., Firtel R. A. A secreted factor and cyclic AMP jointly regulate cell-type-specific gene expression in Dictyostelium discoideum. Mol Cell Biol. 1985 Apr;5(4):705–713. doi: 10.1128/mcb.5.4.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Oohata A. A. A prestalk-cell-specific acid phosphatase in Dictyostelium discoideum. J Embryol Exp Morphol. 1983 Apr;74:311–319. [PubMed] [Google Scholar]
  29. Ratner D., Borth W. Comparison of differentiating Dictyostelium discoideum cell types separated by an improved method of density gradient centrifugation. Exp Cell Res. 1983 Jan;143(1):1–13. doi: 10.1016/0014-4827(83)90103-9. [DOI] [PubMed] [Google Scholar]
  30. Reymond C. D., Gomer R. H., Mehdy M. C., Firtel R. A. Developmental regulation of a Dictyostelium gene encoding a protein homologous to mammalian ras protein. Cell. 1984 Nov;39(1):141–148. doi: 10.1016/0092-8674(84)90199-5. [DOI] [PubMed] [Google Scholar]
  31. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Saxe C. L., 3rd, Firtel R. A. Analysis of gene expression in rapidly developing mutants of Dictyostelium discoideum. Dev Biol. 1986 Jun;115(2):407–414. doi: 10.1016/0012-1606(86)90260-5. [DOI] [PubMed] [Google Scholar]
  33. Staden R. Graphic methods to determine the function of nucleic acid sequences. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):521–538. doi: 10.1093/nar/12.1part2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tasaka M., Noce T., Takeuchi I. Prestalk and prespore differentiation in Dictyostelium as detected by cell type-specific monoclonal antibodies. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5340–5344. doi: 10.1073/pnas.80.17.5340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Teeter M. M., Mazer J. A., L'Italien J. J. Primary structure of the hydrophobic plant protein crambin. Biochemistry. 1981 Sep 15;20(19):5437–5443. doi: 10.1021/bi00522a013. [DOI] [PubMed] [Google Scholar]
  36. Weijer C. J., Durston A. J. Influence of cyclic AMP and hydrolysis products on cell type regulation in Dictyostelium discoideum. J Embryol Exp Morphol. 1985 Apr;86:19–37. [PubMed] [Google Scholar]
  37. Williams J. G., North M. J., Mahbubani H. A developmentally regulated cysteine proteinase in Dictyostelium discoideum. EMBO J. 1985 Apr;4(4):999–1006. doi: 10.1002/j.1460-2075.1985.tb03730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES