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. 1994 Sep;14(9):5840–5849. doi: 10.1128/mcb.14.9.5840

A GBF-binding site and a novel AT element define the minimal sequences sufficient to direct prespore-specific expression in Dictyostelium discoideum.

J A Powell-Coffman 1, G R Schnitzler 1, R A Firtel 1
PMCID: PMC359110  PMID: 8065317

Abstract

In order to better understand the molecular mechanisms of cellular differentiation in Dictyostelium discoideum, we have identified the minimum regulatory sequences of the prespore-specific gene SP60/cotC that are sufficient to confer cell-type-specific expression on a heterologous promoter. This region includes at least two essential cis-acting elements: a novel AT-rich element (or elements) and CAE3. The essential function of the AT element is confirmed through point mutations that decrease expression below the level of detection. CAE3 is one of three CA-rich elements (CAEs) required for the induction of SP60/cotC during development or in response to extracellular cyclic AMP. The CAEs have differential affinities for a specific developmentally induced nuclear activity (CAE1 > CAE2 >> CAE3). Here, we identify this activity as G-box-binding factor (GBF) and show that in vitro-transcribed and -translated GBF binds all three SP60/cotC CAEs in a sequence-specific manner. Previous studies have suggested that GBF mediates the induction of some prestalk genes, and these results demonstrate that it also has a specific role in prespore gene activation.

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Selected References

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  1. Ceccarelli A., Mahbubani H. J., Insall R., Schnitzler G., Firtel R. A., Williams J. G. A G-rich sequence element common to Dictyostelium genes which differ radically in their patterns of expression. Dev Biol. 1992 Jul;152(1):188–193. doi: 10.1016/0012-1606(92)90169-h. [DOI] [PubMed] [Google Scholar]
  2. Ceccarelli A., Mahbubani H., Williams J. G. Positively and negatively acting signals regulating stalk cell and anterior-like cell differentiation in Dictyostelium. Cell. 1991 Jun 14;65(6):983–989. doi: 10.1016/0092-8674(91)90550-i. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Cubitt A. B., Carrel F., Dharmawardhane S., Gaskins C., Hadwiger J., Howard P., Mann S. K., Okaichi K., Zhou K., Firtel R. A. Molecular genetic analysis of signal transduction pathways controlling multicellular development in Dictyostelium. Cold Spring Harb Symp Quant Biol. 1992;57:177–192. doi: 10.1101/sqb.1992.057.01.023. [DOI] [PubMed] [Google Scholar]
  5. Datta S., Firtel R. A. An 80-bp cis-acting regulatory region controls cAMP and development regulation of a prestalk gene in Dictyostelium. Genes Dev. 1988 Mar;2(3):294–304. doi: 10.1101/gad.2.3.294. [DOI] [PubMed] [Google Scholar]
  6. Dingermann T., Reindl N., Werner H., Hildebrandt M., Nellen W., Harwood A., Williams J., Nerke K. Optimization and in situ detection of Escherichia coli beta-galactosidase gene expression in Dictyostelium discoideum. Gene. 1989 Dec 28;85(2):353–362. doi: 10.1016/0378-1119(89)90428-9. [DOI] [PubMed] [Google Scholar]
  7. Early A. E., Gaskell M. J., Traynor D., Williams J. G. Two distinct populations of prestalk cells within the tip of the migratory Dictyostelium slug with differing fates at culmination. Development. 1993 Jun;118(2):353–362. doi: 10.1242/dev.118.2.353. [DOI] [PubMed] [Google Scholar]
  8. Early A. E., Williams J. G. A Dictyostelium prespore-specific gene is transcriptionally repressed by DIF in vitro. Development. 1988 Jul;103(3):519–524. doi: 10.1242/dev.103.3.519. [DOI] [PubMed] [Google Scholar]
  9. Early A. E., Williams J. G. Identification of sequences regulating the transcription of a Dictyostelium gene selectively expressed in prespore cells. Nucleic Acids Res. 1989 Aug 25;17(16):6473–6484. doi: 10.1093/nar/17.16.6473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fosnaugh K. L., Loomis W. F. Coordinate regulation of the spore coat genes in Dictyostelium discoideum. Dev Genet. 1991;12(1-2):123–132. doi: 10.1002/dvg.1020120120. [DOI] [PubMed] [Google Scholar]
  11. Fosnaugh K. L., Loomis W. F. Enhancer regions responsible for temporal and cell-type-specific expression of a spore coat gene in Dictyostelium. Dev Biol. 1993 May;157(1):38–48. doi: 10.1006/dbio.1993.1110. [DOI] [PubMed] [Google Scholar]
  12. Gomer R. H., Datta S., Firtel R. A. 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. J Cell Biol. 1986 Nov;103(5):1999–2015. doi: 10.1083/jcb.103.5.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gomer R. H., Firtel R. A. Cell-autonomous determination of cell-type choice in Dictyostelium development by cell-cycle phase. Science. 1987 Aug 14;237(4816):758–762. doi: 10.1126/science.3039657. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Haberstroh L., Galindo J., Firtel R. A. Developmental and spatial regulation of a Dictyostelium prespore gene: cis-acting elements and a cAMP-induced, developmentally regulated DNA binding activity. Development. 1991 Nov;113(3):947–958. doi: 10.1242/dev.113.3.947. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Hjorth A. L., Khanna N. C., Firtel R. A. A trans-acting factor required for cAMP-induced gene expression in Dictyostelium is regulated developmentally and induced by cAMP. Genes Dev. 1989 Jun;3(6):747–759. doi: 10.1101/gad.3.6.747. [DOI] [PubMed] [Google Scholar]
  18. Hjorth A. L., Pears C., Williams J. G., Firtel R. A. A developmentally regulated trans-acting factor recognizes dissimilar G/C-rich elements controlling a class of cAMP-inducible Dictyostelium genes. Genes Dev. 1990 Mar;4(3):419–432. doi: 10.1101/gad.4.3.419. [DOI] [PubMed] [Google Scholar]
  19. Hjorth A., Datta S., Khanna N. C., Firtel R. A. Analysis of cis and trans elements involved in cAMP-inducible gene expression in Dictyostelium discoideum. Dev Genet. 1988;9(4-5):435–454. doi: 10.1002/dvg.1020090422. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Howard P. K., Ahern K. G., Firtel R. A. Establishment of a transient expression system for Dictyostelium discoideum. Nucleic Acids Res. 1988 Mar 25;16(6):2613–2623. doi: 10.1093/nar/16.6.2613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jones N. Transcriptional regulation by dimerization: two sides to an incestuous relationship. Cell. 1990 Apr 6;61(1):9–11. doi: 10.1016/0092-8674(90)90207-u. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Lee K. A. Dimeric transcription factor families: it takes two to tango but who decides on partners and the venue? J Cell Sci. 1992 Sep;103(Pt 1):9–14. doi: 10.1242/jcs.103.1.9. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. 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]
  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. Ozaki T., Nakao H., Orii H., Morio T., Takeuchi I., Tasaka M. Developmental regulation of transcription of a novel prespore-specific gene (Dp87) in Dictyostelium discoideum. Development. 1993 Apr;117(4):1299–1308. doi: 10.1242/dev.117.4.1299. [DOI] [PubMed] [Google Scholar]
  31. Pears C. J., Williams J. G. Multiple copies of a G-rich element upstream of a cAMP-inducible Dictyostelium gene are necessary but not sufficient for efficient gene expression. Nucleic Acids Res. 1988 Sep 12;16(17):8467–8486. doi: 10.1093/nar/16.17.8467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Reichel R. R., Jacob S. T. Control of gene expression by lipophilic hormones. FASEB J. 1993 Mar;7(5):427–436. doi: 10.1096/fasebj.7.5.8385039. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Schnitzler G. R., Fischer W. H., Firtel R. A. Cloning and characterization of the G-box binding factor, an essential component of the developmental switch between early and late development in Dictyostelium. Genes Dev. 1994 Feb 15;8(4):502–514. doi: 10.1101/gad.8.4.502. [DOI] [PubMed] [Google Scholar]
  35. Tasaka M., Hasegawa M., Nakata M., Orii H., Ozaki T., Takeuchi I. Protein binding and DNase-I-hypersensitive sites in the cis-acting regulatory region of the spore-coat SP96 gene of Dictyostelium. Mech Dev. 1992 Feb;36(3):105–115. doi: 10.1016/0925-4773(92)90062-o. [DOI] [PubMed] [Google Scholar]
  36. Weijer C. J., Duschl G., David C. N. Dependence of cell-type proportioning and sorting on cell cycle phase in Dictyostelium discoideum. J Cell Sci. 1984 Aug;70:133–145. doi: 10.1242/jcs.70.1.133. [DOI] [PubMed] [Google Scholar]
  37. Williams J. G., Duffy K. T., Lane D. P., McRobbie S. J., Harwood A. J., Traynor D., Kay R. R., Jermyn K. A. Origins of the prestalk-prespore pattern in Dictyostelium development. Cell. 1989 Dec 22;59(6):1157–1163. doi: 10.1016/0092-8674(89)90771-x. [DOI] [PubMed] [Google Scholar]

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