Abstract
The Dictyostelium discoideum cell surface antigen PsA is a glycoprotein which first appears in the multicellular stage soon after tip formation and is selectively expressed on prespore cells. The D19 gene encodes an mRNA sequence which is highly enriched in prespore over prestalk cells in the slug stage. We have determined 81 amino acid residues of N-terminal sequence from immunoaffinity-purified PsA protein and shown this sequence to be identical to the predicted sequence of the D19 gene. There are several short repeat elements close to the C terminus, and unequal crossing-over within these is proposed to account for the size polymorphism observed in PsA protein isolated from different D. discoideum strains. The repeats are proline rich and show similarity to the C-terminal region of the D. discoideum cell adhesion molecule, contact sites A. The extreme C terminus, which is also homologous to contact sites A, is characteristic of proteins attached to the plasma membrane via a glycosyl-phosphatidylinositol link. We have marked the PsA gene by insertion of an oligonucleotide encoding an epitope of the human c-myc protein. A construct containing this gene and 990 base pairs of 5'-flanking region directed correct temporal and spatial mRNA accumulation. We found the marked PsA protein, detected with the human c-myc antibody, to be correctly localized on the surface of cells.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- Barklis E., Pontius B., Barfield K., Lodish H. F. Structure of the promoter of the Dictyostelium discoideum prespore EB4 gene. Mol Cell Biol. 1985 Jun;5(6):1465–1472. doi: 10.1128/mcb.5.6.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bartles J. R., Frazier W. A., Rosen S. D. Slime mold lectins. Int Rev Cytol. 1982;75:61–99. doi: 10.1016/s0074-7696(08)61002-5. [DOI] [PubMed] [Google Scholar]
- Bause E. Structural requirements of N-glycosylation of proteins. Studies with proline peptides as conformational probes. Biochem J. 1983 Feb 1;209(2):331–336. doi: 10.1042/bj2090331. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Briggs M. S., Gierasch L. M. Molecular mechanisms of protein secretion: the role of the signal sequence. Adv Protein Chem. 1986;38:109–180. doi: 10.1016/s0065-3233(08)60527-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Early A. E., Williams J. G. Two vectors which facilitate gene manipulation and a simplified transformation procedure for Dictyostelium discoideum. Gene. 1987;59(1):99–106. doi: 10.1016/0378-1119(87)90270-8. [DOI] [PubMed] [Google Scholar]
- Erdos G. W., Raper K. B., Vogen L. K. Mating Types and Macrocyst Formation in Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1828–1830. doi: 10.1073/pnas.70.6.1828. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Grant W. N., Welker D. L., Williams K. L. A polymorphic, prespore-specific cell surface glycoprotein is present in the extracellular matrix of Dictyostelium discoideum. Mol Cell Biol. 1985 Oct;5(10):2559–2566. doi: 10.1128/mcb.5.10.2559. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hemperly J. J., Edelman G. M., Cunningham B. A. cDNA clones of the neural cell adhesion molecule (N-CAM) lacking a membrane-spanning region consistent with evidence for membrane attachment via a phosphatidylinositol intermediate. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9822–9826. doi: 10.1073/pnas.83.24.9822. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hunkapiller M. W., Hewick R. M., Dreyer W. J., Hood L. E. High-sensitivity sequencing with a gas-phase sequenator. Methods Enzymol. 1983;91:399–413. doi: 10.1016/s0076-6879(83)91038-8. [DOI] [PubMed] [Google Scholar]
- Jermyn K. A., Berks M., Kay R. R., Williams J. G. Two distinct classes of prestalk-enriched mRNA sequences in Dictyostelium discoideum. Development. 1987 Aug;100(4):745–755. doi: 10.1242/dev.100.4.745. [DOI] [PubMed] [Google Scholar]
- Krefft M., Voet L., Gregg J. H., Mairhofer H., Williams K. L. Evidence that positional information is used to establish the prestalk-prespore pattern in Dictyostelium discoideum aggregates. EMBO J. 1984 Jan;3(1):201–206. doi: 10.1002/j.1460-2075.1984.tb01784.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krefft M., Voet L., Mairhofer H., Williams K. L. Analysis of proportion regulation in slugs of Dictyostelium discoideum using a monoclonal antibody and a FACS-IV. Exp Cell Res. 1983 Aug;147(1):235–239. doi: 10.1016/0014-4827(83)90291-4. [DOI] [PubMed] [Google Scholar]
- Kyhse-Andersen J. Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods. 1984 Dec;10(3-4):203–209. doi: 10.1016/0165-022x(84)90040-x. [DOI] [PubMed] [Google Scholar]
- Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Mangiarotti G., Ceccarelli A., Lodish H. F. Cyclic AMP stabilizes a class of developmentally regulated Dictyostelium discoideum mRNAs. Nature. 1983 Feb 17;301(5901):616–618. doi: 10.1038/301616a0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Munro S., Pelham H. R. An Hsp70-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein. Cell. 1986 Jul 18;46(2):291–300. doi: 10.1016/0092-8674(86)90746-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Oyama M., Blumberg D. D. Changes during differentiation in requirements for cAMP for expression of cell-type-specific mRNAs in the cellular slime mold, Dictyostelium discoideum. Dev Biol. 1986 Oct;117(2):550–556. doi: 10.1016/0012-1606(86)90323-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Sadeghi H., Williams K., Klein C. Characterization of antigen 117. A developmentally regulated cell surface glycoprotein of Dictyostelium discoideum. J Biol Chem. 1987 Dec 5;262(34):16294–16299. [PubMed] [Google Scholar]
- Schaap P., Van Lookeren Campagne M. M., Van Driel R., Spek W., Van Haastert P. J., Pinas J. Postaggregative differentiation induction by cyclic AMP in Dictyostelium: intracellular transduction pathway and requirement for additional stimuli. Dev Biol. 1986 Nov;118(1):52–63. doi: 10.1016/0012-1606(86)90072-2. [DOI] [PubMed] [Google Scholar]
- Schaap P., Wang M. Interactions between adenosine and oscillatory cAMP signaling regulate size and pattern in Dictyostelium. Cell. 1986 Apr 11;45(1):137–144. doi: 10.1016/0092-8674(86)90545-3. [DOI] [PubMed] [Google Scholar]
- Smith E., Roberts K., Butcher G. W., Galfre G. Monoclonal antibody screening: two methods using antigens immobilized on nitrocellulose. Anal Biochem. 1984 Apr;138(1):119–124. doi: 10.1016/0003-2697(84)90778-4. [DOI] [PubMed] [Google Scholar]
- Standring R., Williams A. F. Glycoproteins and antigens of membranes prepared from rat thymocytes after lysis by shearing or with the detergent Tween-40. Biochim Biophys Acta. 1978 Mar 21;508(1):85–96. doi: 10.1016/0005-2736(78)90190-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Tsang A., Bradbury J. M. Separation and properties of prestalk and prespore cells of Dictyostelium discoideum. Exp Cell Res. 1981 Apr;132(2):433–441. doi: 10.1016/0014-4827(81)90118-x. [DOI] [PubMed] [Google Scholar]
- Watts D. J., Ashworth J. M. Growth of myxameobae of the cellular slime mould Dictyostelium discoideum in axenic culture. Biochem J. 1970 Sep;119(2):171–174. doi: 10.1042/bj1190171. [DOI] [PMC free article] [PubMed] [Google Scholar]
- West C. M., Loomis W. F. Absence of a carbohydrate modification does not affect the level or subcellular localization of three membrane glycoproteins in modB mutants of Dictyostelium discoideum. J Biol Chem. 1985 Nov 5;260(25):13803–13809. [PubMed] [Google Scholar]