Abstract
The activity of cAMP-dependent protein kinase (PKA) is required for proper development at several stages during the Dictyostelium life cycle. We present evidence that activation of PKA is rate-limiting for the differentiation of prespore cells to spores and that PKA activation may be the developmental trigger for sporulation. Strains that overexpress the gene encoding the catalytic subunit of PKA (PKAcat) or lack a functional regulatory subunit (rdeC strains) undergo rapid, heterochronic development. We show that overexpression of PKAcat in prespore cell is sufficient to directly induce expression of the spore maturation marker spiA and differentiation to spores, in a cell-autonomous manner. Moreover, overexpression of PKAcat in prespore cells can bypass a mutation that blocks an earlier developmental step to induce spiA expression. Our results suggest that the regulatory pathway in prespore cells between the activation of PKA and spiA induction/spore maturation is quite short and that PKAcat expression in prespore cells may mediate spore differentiation at the level of transcription. This induction of sporulation requires the prior activation of the prespore cell pathway. In addition, we show that beta-galactosidase activity expressed from a PKAcat promoter/lacZ reporter construct is highly enriched in the anterior prestalk A region during the tipped aggregate, slug, and early culminant stages and that this pattern switches abruptly to a prespore pattern at the time of spore maturation, supporting the proposed role of PKA in this process.
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- Abe K., Yanagisawa K. A new class of rapidly developing mutants in Dictyostelium discoideum: implications for cyclic AMP metabolism and cell differentiation. Dev Biol. 1983 Jan;95(1):200–210. doi: 10.1016/0012-1606(83)90018-0. [DOI] [PubMed] [Google Scholar]
- Anjard C., Pinaud S., Kay R. R., Reymond C. D. Overexpression of Dd PK2 protein kinase causes rapid development and affects the intracellular cAMP pathway of Dictyostelium discoideum. Development. 1992 Jul;115(3):785–790. doi: 10.1242/dev.115.3.785. [DOI] [PubMed] [Google Scholar]
- Bürki E., Anjard C., Scholder J. C., Reymond C. D. Isolation of two genes encoding putative protein kinases regulated during Dictyostelium discoideum development. Gene. 1991 Jun 15;102(1):57–65. doi: 10.1016/0378-1119(91)90538-m. [DOI] [PubMed] [Google Scholar]
- Devreotes P. Dictyostelium discoideum: a model system for cell-cell interactions in development. Science. 1989 Sep 8;245(4922):1054–1058. doi: 10.1126/science.2672337. [DOI] [PubMed] [Google Scholar]
- Esch R. K., Firtel R. A. cAMP and cell sorting control the spatial expression of a developmentally essential cell-type-specific ras gene in Dictyostelium. Genes Dev. 1991 Jan;5(1):9–21. doi: 10.1101/gad.5.1.9. [DOI] [PubMed] [Google Scholar]
- Firtel R. A., Chapman A. L. A role for cAMP-dependent protein kinase A in early Dictyostelium development. Genes Dev. 1990 Jan;4(1):18–28. doi: 10.1101/gad.4.1.18. [DOI] [PubMed] [Google Scholar]
- Firtel R. A. Signal transduction pathways controlling multicellular development in Dictyostelium. Trends Genet. 1991 Nov-Dec;7(11-12):381–388. doi: 10.1016/0168-9525(91)90260-w. [DOI] [PubMed] [Google Scholar]
- Gerisch G. Cyclic AMP and other signals controlling cell development and differentiation in Dictyostelium. Annu Rev Biochem. 1987;56:853–879. doi: 10.1146/annurev.bi.56.070187.004225. [DOI] [PubMed] [Google Scholar]
- Gunzburg J., Veron M. A cAMP-dependent protein kinase is present in differentiating Dictyostelium discoideum cells. EMBO J. 1982;1(9):1063–1068. doi: 10.1002/j.1460-2075.1982.tb01297.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haberstroh L., Firtel R. A. A spatial gradient of expression of a cAMP-regulated prespore cell-type-specific gene in Dictyostelium. Genes Dev. 1990 Apr;4(4):596–612. doi: 10.1101/gad.4.4.596. [DOI] [PubMed] [Google Scholar]
- Hadwiger J. A., Firtel R. A. Analysis of G alpha 4, a G-protein subunit required for multicellular development in Dictyostelium. Genes Dev. 1992 Jan;6(1):38–49. doi: 10.1101/gad.6.1.38. [DOI] [PubMed] [Google Scholar]
- Harwood A. J., Hopper N. A., Simon M. N., Bouzid S., Veron M., Williams J. G. Multiple roles for cAMP-dependent protein kinase during Dictyostelium development. Dev Biol. 1992 Jan;149(1):90–99. doi: 10.1016/0012-1606(92)90266-j. [DOI] [PubMed] [Google Scholar]
- Harwood A. J., Hopper N. A., Simon M. N., Driscoll D. M., Veron M., Williams J. G. Culmination in Dictyostelium is regulated by the cAMP-dependent protein kinase. Cell. 1992 May 15;69(4):615–624. doi: 10.1016/0092-8674(92)90225-2. [DOI] [PubMed] [Google Scholar]
- Hopper N. A., Anjard C., Reymond C. D., Williams J. G. Induction of terminal differentiation of Dictyostelium by cAMP-dependent protein kinase and opposing effects of intracellulr and extracellular cAMP on stalk cell differentiation. Development. 1993 Sep;119(1):147–154. doi: 10.1242/dev.119.1.147. [DOI] [PubMed] [Google Scholar]
- Hopper N. A., Harwood A. J., Bouzid S., Véron M., Williams J. G. Activation of the prespore and spore cell pathway of Dictyostelium differentiation by cAMP-dependent protein kinase and evidence for its upstream regulation by ammonia. EMBO J. 1993 Jun;12(6):2459–2466. doi: 10.1002/j.1460-2075.1993.tb05900.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howard P. K., Sefton B. M., Firtel R. A. Analysis of a spatially regulated phosphotyrosine phosphatase identifies tyrosine phosphorylation as a key regulatory pathway in Dictyostelium. Cell. 1992 Nov 13;71(4):637–647. doi: 10.1016/0092-8674(92)90597-6. [DOI] [PubMed] [Google Scholar]
- Kimmel A. R., Firtel R. A. cAMP signal transduction pathways regulating development of Dictyostelium discoideum. Curr Opin Genet Dev. 1991 Oct;1(3):383–390. doi: 10.1016/s0959-437x(05)80304-1. [DOI] [PubMed] [Google Scholar]
- Majerfeld I. H., Leichtling B. H., Meligeni J. A., Spitz E., Rickenberg H. V. A cytosolic cyclic AMP-dependent protein kinase in Dictyostelium discoideum. I. Properties. J Biol Chem. 1984 Jan 10;259(1):654–661. [PubMed] [Google Scholar]
- Mann S. K., Firtel R. A. A developmentally regulated, putative serine/threonine protein kinase is essential for development in Dictyostelium. Mech Dev. 1991 Sep;35(2):89–101. doi: 10.1016/0925-4773(91)90060-j. [DOI] [PubMed] [Google Scholar]
- Mann S. K., Firtel R. A. Cyclic AMP regulation of early gene expression in Dictyostelium discoideum: mediation via the cell surface cyclic AMP receptor. Mol Cell Biol. 1987 Jan;7(1):458–469. doi: 10.1128/mcb.7.1.458. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mann S. K., Firtel R. A. cAMP-dependent protein kinase differentially regulates prestalk and prespore differentiation during Dictyostelium development. Development. 1993 Sep;119(1):135–146. doi: 10.1242/dev.119.1.135. [DOI] [PubMed] [Google Scholar]
- Mann S. K., Yonemoto W. M., Taylor S. S., Firtel R. A. DdPK3, which plays essential roles during Dictyostelium development, encodes the catalytic subunit of cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10701–10705. doi: 10.1073/pnas.89.22.10701. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pitt G. S., Brandt R., Lin K. C., Devreotes P. N., Schaap P. Extracellular cAMP is sufficient to restore developmental gene expression and morphogenesis in Dictyostelium cells lacking the aggregation adenylyl cyclase (ACA). Genes Dev. 1993 Nov;7(11):2172–2180. doi: 10.1101/gad.7.11.2172. [DOI] [PubMed] [Google Scholar]
- Richardson D. L., Hong C. B., Loomis W. F. A prespore gene, Dd31, expressed during culmination of Dictyostelium discoideum. Dev Biol. 1991 Apr;144(2):269–280. doi: 10.1016/0012-1606(91)90421-x. [DOI] [PubMed] [Google Scholar]
- Richardson D. L., Loomis W. F. Disruption of the sporulation-specific gene spiA in Dictyostelium discoideum leads to spore instability. Genes Dev. 1992 Jun;6(6):1058–1070. doi: 10.1101/gad.6.6.1058. [DOI] [PubMed] [Google Scholar]
- Riley B. B., Jensen B. R., Barclay S. L. Conditions that elevate intracellular cyclic AMP levels promote spore formation in Dictyostelium. Differentiation. 1989 Jul;41(1):5–13. doi: 10.1111/j.1432-0436.1989.tb00726.x. [DOI] [PubMed] [Google Scholar]
- Rutherford C. L., Taylor R. D., Frame L. T., Auck R. L. A cyclic AMP dependent protein kinase in Dictyostelium discoideum. Biochem Biophys Res Commun. 1982 Oct 15;108(3):1210–1220. doi: 10.1016/0006-291x(82)92129-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Simon M. N., Pelegrini O., Veron M., Kay R. R. Mutation of protein kinase A causes heterochronic development of Dictyostelium. Nature. 1992 Mar 12;356(6365):171–172. doi: 10.1038/356171a0. [DOI] [PubMed] [Google Scholar]
- Van Haastert P. J. Transmembrane signal transduction pathways in Dictyostelium. Adv Second Messenger Phosphoprotein Res. 1991;23:185–226. [PubMed] [Google Scholar]
- Williams J. G. Regulation of cellular differentiation during Dictyostelium morphogenesis. Curr Opin Genet Dev. 1991 Oct;1(3):358–362. doi: 10.1016/s0959-437x(05)80300-4. [DOI] [PubMed] [Google Scholar]