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. 1992 Jun 11;20(11):2795–2802. doi: 10.1093/nar/20.11.2795

A negative transcriptional control region of a developmentally-regulated gene co-localizes with the origin of replication of an endogenous plasmid in Dictyostelium.

J A Powell 1, J Galindo 1, R A Firtel 1
PMCID: PMC336924  PMID: 1319575

Abstract

The endogenous nuclear plasmid Ddp1 from the wild-type Dictyostelium discoideum strain NC4 has been cloned, its origin of replication has been localized, and plasmid-encoded genes have been mapped that are preferentially expressed during growth or development. Here we present an analysis of the regulation of the Ddp1-encoded gene d5, which, in wild-type cells, is expressed only during the multicellular stages of development. In this study, we show that sequences 3' to the d5 coding region are required to suppress constitutive expression of d5 from aberrant transcriptional start sites and that this regulatory region acts at a distance and in an orientation-independent manner. The cis-acting negative regulatory element(s) necessary for repression of aberrant d5 expression is either very tightly linked or identical to sequences required for extrachromosomal replication, such that all 3' deletions that cause constitutive d5 expression result in the integration of the plasmid into the D. discoideum genome. Placing d5 (without the 3' regions containing the Ddp1 origin) on an extrachromosomal vector based on another endogenous plasmid (Ddp2) did not restore proper transcriptional regulation, suggesting that an extrachromosomal environment alone is not sufficient to confer proper transcriptional regulation to d5.

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Selected References

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

  1. Ahern K. G., Howard P. K., Firtel R. A. Identification of regions essential for extrachromosomal replication and maintenance of an endogenous plasmid in Dictyostelium. Nucleic Acids Res. 1988 Jul 25;16(14B):6825–6837. doi: 10.1093/nar/16.14.6825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ariga H., Imamura Y., Iguchi-Ariga S. M. DNA replication origin and transcriptional enhancer in c-myc gene share the c-myc protein binding sequences. EMBO J. 1989 Dec 20;8(13):4273–4279. doi: 10.1002/j.1460-2075.1989.tb08613.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ariizumi K., Takahashi H., Nakamura M., Ariga H. Effect of silencer on polyomavirus DNA replication. Mol Cell Biol. 1989 Sep;9(9):4026–4031. doi: 10.1128/mcb.9.9.4026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ashktorab H., Welker D. L. Establishment of the nuclear location of Dictyostelium discoideum plasmids. Gene. 1988 May 15;65(1):41–49. doi: 10.1016/0378-1119(88)90415-5. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Brand A. H., Micklem G., Nasmyth K. A yeast silencer contains sequences that can promote autonomous plasmid replication and transcriptional activation. Cell. 1987 Dec 4;51(5):709–719. doi: 10.1016/0092-8674(87)90094-8. [DOI] [PubMed] [Google Scholar]
  7. Calame K. L. Immunoglobulin gene transcription: molecular mechanisms. Trends Genet. 1989 Dec;5(12):395–399. doi: 10.1016/0168-9525(89)90197-2. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Datta S., Firtel R. A. Identification of the sequences controlling cyclic AMP regulation and cell-type-specific expression of a prestalk-specific gene in Dictyostelium discoideum. Mol Cell Biol. 1987 Jan;7(1):149–159. doi: 10.1128/mcb.7.1.149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DeLucia A. L., Deb S., Partin K., Tegtmeyer P. Functional interactions of the simian virus 40 core origin of replication with flanking regulatory sequences. J Virol. 1986 Jan;57(1):138–144. doi: 10.1128/jvi.57.1.138-144.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Dynes J. L., Firtel R. A. Molecular complementation of a genetic marker in Dictyostelium using a genomic DNA library. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7966–7970. doi: 10.1073/pnas.86.20.7966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Emerson B. M., Nickol J. M., Jackson P. D., Felsenfeld G. Analysis of the tissue-specific enhancer at the 3' end of the chicken adult beta-globin gene. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4786–4790. doi: 10.1073/pnas.84.14.4786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Farrar N. A., Longhurst T. J., Leigh D. A., Williams K. L. Nucleotide sequence of a developmentally regulated Dictyostelium discoideum endogenous plasmid gene and 5' flanking region. Nucleic Acids Res. 1988 Nov 25;16(22):10914–10914. doi: 10.1093/nar/16.22.10914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Farrar N. A., Williams K. L. Nuclear plasmids in the simple eukaryotes Saccharomyces cerevisiae and Dictyostelium discoideum. Trends Genet. 1988 Dec;4(12):343–348. doi: 10.1016/0168-9525(88)90054-6. [DOI] [PubMed] [Google Scholar]
  16. Firtel R. A., Silan C., Ward T. E., Howard P., Metz B. A., Nellen W., Jacobson A. Extrachromosomal replication of shuttle vectors in Dictyostelium discoideum. Mol Cell Biol. 1985 Nov;5(11):3241–3250. doi: 10.1128/mcb.5.11.3241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Firtel R. A., van Haastert P. J., Kimmel A. R., Devreotes P. N. G protein linked signal transduction pathways in development: dictyostelium as an experimental system. Cell. 1989 Jul 28;58(2):235–239. doi: 10.1016/0092-8674(89)90837-4. [DOI] [PubMed] [Google Scholar]
  18. Georgopoulos K., van den Elsen P., Bier E., Maxam A., Terhorst C. A T cell-specific enhancer is located in a DNase I-hypersensitive area at the 3' end of the CD3-delta gene. EMBO J. 1988 Aug;7(8):2401–2407. doi: 10.1002/j.1460-2075.1988.tb03085.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gurniak C. B., Bang A. G., Noegel A. A. Transcript and sequence analysis of a 5.1 kb contiguous fragment of Dictyostelium discoideum plasmid Ddp1 that contains the origin of replication and codes for several transcripts. Curr Genet. 1990 Apr;17(4):321–325. doi: 10.1007/BF00314879. [DOI] [PubMed] [Google Scholar]
  20. 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]
  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. Hughes J. E., Ashtorab H., Welker D. L. Nuclear plasmids in the Dictyostelium slime molds. Dev Genet. 1988;9(4-5):495–504. doi: 10.1002/dvg.1020090426. [DOI] [PubMed] [Google Scholar]
  23. Kimmerly W., Buchman A., Kornberg R., Rine J. Roles of two DNA-binding factors in replication, segregation and transcriptional repression mediated by a yeast silencer. EMBO J. 1988 Jul;7(7):2241–2253. doi: 10.1002/j.1460-2075.1988.tb03064.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klein P. S., Sun T. J., Saxe C. L., 3rd, Kimmel A. R., Johnson R. L., Devreotes P. N. A chemoattractant receptor controls development in Dictyostelium discoideum. Science. 1988 Sep 16;241(4872):1467–1472. doi: 10.1126/science.3047871. [DOI] [PubMed] [Google Scholar]
  25. Kumagai A., Pupillo M., Gundersen R., Miake-Lye R., Devreotes P. N., Firtel R. A. Regulation and function of G alpha protein subunits in Dictyostelium. Cell. 1989 Apr 21;57(2):265–275. doi: 10.1016/0092-8674(89)90964-1. [DOI] [PubMed] [Google Scholar]
  26. Leiting B., Lindner I. J., Noegel A. A. The extrachromosomal replication of Dictyostelium plasmid Ddp2 requires a cis-acting element and a plasmid-encoded trans-acting factor. Mol Cell Biol. 1990 Jul;10(7):3727–3736. doi: 10.1128/mcb.10.7.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Leiting B., Noegel A. Construction of an extrachromosomally replicating transformation vector for Dictyostelium discoideum. Plasmid. 1988 Nov;20(3):241–248. doi: 10.1016/0147-619x(88)90030-3. [DOI] [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. Nellen W., Datta S., Reymond C., Sivertsen A., Mann S., Crowley T., Firtel R. A. Molecular biology in Dictyostelium: tools and applications. Methods Cell Biol. 1987;28:67–100. doi: 10.1016/s0091-679x(08)61637-4. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Noegel A., Metz B. A., Williams K. L. Developmentally regulated transcription of Dictyostelium discoideum plasmid Ddp1. EMBO J. 1985 Dec 30;4(13B):3797–3803. doi: 10.1002/j.1460-2075.1985.tb04150.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Noegel A., Welker D. L., Metz B. A., Williams K. L. Presence of nuclear associated plasmids in the lower eukaryote Dictyostelium discoideum. J Mol Biol. 1985 Sep 20;185(2):447–450. doi: 10.1016/0022-2836(85)90416-4. [DOI] [PubMed] [Google Scholar]
  34. Orii H., Suzuki K., Tanaka Y., Yanagisawa K. A new type of plasmid from a wild isolate of Dictyostelium species: the existence of closely situated long inverted repeats. Nucleic Acids Res. 1987 Feb 11;15(3):1097–1107. doi: 10.1093/nar/15.3.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Perez-Stable C., Costantini F. Roles of fetal G gamma-globin promoter elements and the adult beta-globin 3' enhancer in the stage-specific expression of globin genes. Mol Cell Biol. 1990 Mar;10(3):1116–1125. doi: 10.1128/mcb.10.3.1116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ramirez F., Di Liberto M. Complex and diversified regulatory programs control the expression of vertebrate collagen genes. FASEB J. 1990 Apr 1;4(6):1616–1623. doi: 10.1096/fasebj.4.6.2180769. [DOI] [PubMed] [Google Scholar]
  38. Stenlund A., Bream G. L., Botchan M. R. A promoter with an internal regulatory domain is part of the origin of replication in BPV-1. Science. 1987 Jun 26;236(4809):1666–1671. doi: 10.1126/science.3037693. [DOI] [PubMed] [Google Scholar]

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