Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Dec 11;20(23):6235–6238. doi: 10.1093/nar/20.23.6235

Transcriptional regulation by folate: inducible gene expression in Dictyostelium transformants during growth and early development.

J Blusch 1, P Morandini 1, W Nellen 1
PMCID: PMC334510  PMID: 1475184

Abstract

The Dictyostelium discoidin genes are induced in bacteria-grown cells shortly before the onset of development but are also highly expressed during growth in axenic medium. We here show that axenically growing cells strongly respond to the extracellular signal folate by suppressing discoidin synthesis while cell growth and development is not substantially affected. Repression occurs via two previously identified promoter elements, the dIE and the dAXE. Removal of the signal molecules or setting cells up for development results in rapid reactivation of the promoter. Based on this observation, we constructed the transformation vector pVEII and describe a convenient method which allows for controlled expression of a gene of interest in growing cells and also for external modulation in early development. Deletion constructs of the discoidin promoter can be used in addition to vary transcriptional activity over about one order of magnitude.

Full text

PDF
6238

Images in this article

Selected References

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

  1. Ceccarelli A., McRobbie S. J., Jermyn K. A., Duffy K., Early A., Williams J. G. Structural and functional characterization of a Dictyostelium gene encoding a DIF inducible, prestalk-enriched mRNA sequence. Nucleic Acids Res. 1987 Sep 25;15(18):7463–7476. doi: 10.1093/nar/15.18.7463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clarke M., Kayman S. C., Riley K. Density-dependent induction of discoidin-I synthesis in exponentially growing cells of Dictyostelium discoideum. Differentiation. 1987;34(2):79–87. doi: 10.1111/j.1432-0436.1987.tb00053.x. [DOI] [PubMed] [Google Scholar]
  3. Clarke M., Yang J., Kayman S. C. Analysis of the prestarvation response in growing cells of Dictyostelium discoideum. Dev Genet. 1988;9(4-5):315–326. doi: 10.1002/dvg.1020090413. [DOI] [PubMed] [Google Scholar]
  4. Crowley T. E., Nellen W., Gomer R. H., Firtel R. A. Phenocopy of discoidin I-minus mutants by antisense transformation in Dictyostelium. Cell. 1985 Dec;43(3 Pt 2):633–641. doi: 10.1016/0092-8674(85)90235-1. [DOI] [PubMed] [Google Scholar]
  5. Devine J. M., Tsang A. S., Williams J. G. Differential expression of the members of the discoidin I multigene family during growth and development of Dictyostelium discoideum. Cell. 1982 Apr;28(4):793–800. doi: 10.1016/0092-8674(82)90058-7. [DOI] [PubMed] [Google Scholar]
  6. Dingermann T., Troidl E. M., Bröker M., Nerke K. Expression of human antithrombin III in the cellular slime mould Dictyostelium discoideum. Appl Microbiol Biotechnol. 1991 Jul;35(4):496–503. doi: 10.1007/BF00169756. [DOI] [PubMed] [Google Scholar]
  7. Dingermann T., Werner H., Schütz A., Zündorf I., Nerke K., Knecht D., Marschalek R. Establishment of a system for conditional gene expression using an inducible tRNA suppressor gene. Mol Cell Biol. 1992 Sep;12(9):4038–4045. doi: 10.1128/mcb.12.9.4038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Faix J., Gerisch G., Noegel A. A. Constitutive overexpression of the contact site A glycoprotein enables growth-phase cells of Dictyostelium discoideum to aggregate. EMBO J. 1990 Sep;9(9):2709–2716. doi: 10.1002/j.1460-2075.1990.tb07457.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Faix J., Gerisch G., Noegel A. A. Overexpression of the csA cell adhesion molecule under its own cAMP-regulated promoter impairs morphogenesis in Dictyostelium. J Cell Sci. 1992 Jun;102(Pt 2):203–214. doi: 10.1242/jcs.102.2.203. [DOI] [PubMed] [Google Scholar]
  10. Fasel N., Begdadi-Rais C., Bernard M., Bron C., Corradin G., Reymond C. D. Dictyostelium discoideum as an expression host for the circumsporozoite protein of Plasmodium falciparum. Gene. 1992 Feb 15;111(2):157–163. doi: 10.1016/0378-1119(92)90683-g. [DOI] [PubMed] [Google Scholar]
  11. Hildebrandt M., Humbel B. M., Nellen W. The Dictyostelium discoideum EB4 gene product and a truncated mutant form of the protein are localized in prespore vesicles but absent from mature spores. Dev Biol. 1991 Mar;144(1):212–214. doi: 10.1016/0012-1606(91)90492-l. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Knecht D. A., Loomis W. F. Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum. Science. 1987 May 29;236(4805):1081–1086. doi: 10.1126/science.3576221. [DOI] [PubMed] [Google Scholar]
  14. Maniak M., Saur U., Nellen W. A colony-blot technique for the detection of specific transcripts in eukaryotes. Anal Biochem. 1989 Jan;176(1):78–81. doi: 10.1016/0003-2697(89)90275-3. [DOI] [PubMed] [Google Scholar]
  15. May T., Blusch J., Sachse A., Nellen W. A cis-acting element responsible for early gene induction by extracellular cAMP in Dictyostelium discoideum. Mech Dev. 1991 Feb;33(2):147–155. doi: 10.1016/0925-4773(91)90081-g. [DOI] [PubMed] [Google Scholar]
  16. May T., Kern H., Müller-Taubenberger A., Nellen W. Identification of a cis-acting element controlling induction of early gene expression in Dictyostelium discoideum. Mol Cell Biol. 1989 Nov;9(11):4653–4659. doi: 10.1128/mcb.9.11.4653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nellen W., Firtel R. A. High-copy-number transformants and co-transformation in Dictyostelium. Gene. 1985;39(2-3):155–163. doi: 10.1016/0378-1119(85)90309-9. [DOI] [PubMed] [Google Scholar]
  18. Poole S. J., Firtel R. A. Conserved structural features are found upstream from the three co-ordinately regulated discoidin I genes of Dictyostelium discoideum. J Mol Biol. 1984 Jan 15;172(2):203–220. doi: 10.1016/s0022-2836(84)80038-8. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Vauti F., Morandini P., Blusch J., Sachse A., Nellen W. Regulation of the Discoidin I gamma gene in Dictyostelium discoideum: identification of individual promoter elements mediating induction of transcription and repression by cyclic AMP. Mol Cell Biol. 1990 Aug;10(8):4080–4088. doi: 10.1128/mcb.10.8.4080. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES