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Conserved function in Nicotiana tabacum of a single Drosophila hsp70 promoter heat shock element when fused to a minimal T-DNA promoter

Summary

To demonstrate the extent of evolutionary conservation in the mechanism of induction of heat shock genes between plants and animals, the minimal sequence from the Drosophila hsp70 promoter sufficient to confer heat shock inducible transcription in tobacco was determined. Segments of the hsp70 promoter were fused to a minimal promoter of the T-DNA indole-3-acetamide hydrolase (iaaH) gene, in a chimaeric gene fusion to a neomycin phosphotransferase (NPT II) reporter gene. Sequences bearing one or more heat shock elements (HSEs) rendered the minimal promoter heat shock inducible, with a 37 bp fragment containing a single complete HSE sufficing. The induced NPT II mRNA peaked during the heat shock period, but the maximal level of NPT 11 activity was not observed until 4 h later in the recovery phase, showing that the translation of the NPT II mRNA was shifted from the heat shock period of the recovery phase. That similar sequences containing a single HSE of the Drosophila hsp70 promoter could function in both flies and tobacco indicates the high degree of homology between the two heat shock gene induction systems.

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References

  • Ashburner M, Bonner JJ (1979) The induction of gene activity in Drosophila by heat shock. Cell 17:241–254

    Google Scholar 

  • Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acidcellulose. Proc Natl Acad Sci USA 69:1408–1412

    Google Scholar 

  • Barnett T, Altschuler M, McDaniel CN, Mascarenhas J (1980) Heat shock induced proteins in plant cells. Dev Genet 1:331–340

    Google Scholar 

  • Baumann G, Raschke E, Bevan M, Schöffl F (1987) Functional analysis of sequences required for transcriptional activation of a soybean heat shock gene in transgenic tobacco plants. EMBO J 6:1161–1166

    Google Scholar 

  • Bienz M, Pelham HRB (1986) Heat shock regulatory elements function as an inducible enhancer in the Xenopus hsp70 gene and when linked to a heterologous promoter. Cell 45:753–760

    Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299

    CAS  PubMed  Google Scholar 

  • Cohen RS, Meselson M (1988) Periodic interactions of heat shock transcriptional elements. Nature 332:856–858

    Google Scholar 

  • Czarnecka E, Gurley WB, Nagao RT, Mosquera LA, Key JL (1985) DNA sequence and transcript mapping of a soybean gene encoding a small heat shock protein. Proc Natl Acad Sci USA 82:3726–3730

    Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21

    CAS  Google Scholar 

  • De Vos G, De Beuckeleer M, Van Montagu M, Schell J (1981) Restriction endonuclease mapping of the octopine tumor-inducing plasmid pTiAch5 of Agrobacterium tumefaciens. Plasmid 6:249–253

    Google Scholar 

  • Dudler R, Travers AA (1984) Upstream elements necessary for optimal function of the hsp70 in transformed flies. Cell 38:391–398

    Google Scholar 

  • Feinberg AP, Vogelstein B (1984) A technique for radiolabeling DNA restriction endonculease fragments to high specific activity. Anal Biochem 137:266–267

    CAS  PubMed  Google Scholar 

  • Gielen J, De Beuckeleer M, Seurinck J, Deboeck F, DeGreve H, Lemmers M, Van Montagu M, Schell J (1984) The complete nucleotide sequence of the TL-DNA of the Agrobacterium tumefaciens plasmid pTiAch5. EMBO J 3:835–846

    Google Scholar 

  • Gritz L, Davies J (1983) Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene 25:179–188

    Google Scholar 

  • Hanahan D (1983) Studies on transformations of Escherichia coli with plasmids. J Mol Biol 166:557–580

    CAS  PubMed  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    CAS  Google Scholar 

  • Key JL, Lin CY, Chen YM (1981) Heat shock proteins of higher plants. Proc Natl Acad Sci USA 78:3526–3530

    Google Scholar 

  • Klemenz R, Hultmark D, Gehring WJ (1985) Selective translation of heat shock mRNA in Drosophila melanogaster depends on sequence information in the leader. EMBO J 4:2053–2060

    Google Scholar 

  • Koncz Cs, Schell J (1986) The promoter of T l -DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type of Agrobacterium binary vector. Mol Gen Genet 204:383–396

    CAS  Google Scholar 

  • Lindquist S (1981) Regulation of protein synthesis during heat shock. Nature 293:311–314

    Google Scholar 

  • Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plantarum 18:100–127

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • McKenzie SL, Henikoff S, Meselson M (1975) Localization of RNA from heat-induced polysomes at puff sites in Drosophila melanogaster. Proc Natl Acad Sci USA 72:1117–1121

    Google Scholar 

  • Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Mirault M-E, Southgate R, Delwart E (1982) Regulation of heatshock genes: a DNA sequence upstream of Drosophila hsp70 genes is essential for their induction in monkey cells. EMBO J 1:1279–1285

    CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plantarum 15:473–497

    Google Scholar 

  • Nagao RT, Kimpel JA, Vierling E, Key JL (1986) The heat shock response: a comparative analysis. Oxford Surv Plant Mol Cell Biol 3:384–438

    Google Scholar 

  • Pelham H (1982) A regulatory upstream promoter element in the Drosophila hsp70 heat-shock gene. Cell 30:517–528

    Google Scholar 

  • Pelham H (1985) Activation of heat-shock genes in eukaryotes. Trends Genet 3:31–35

    Google Scholar 

  • Pelham H, Bienz M (1982) A synthetic heat-shock promoter element confers heat-inducibility on the herpes simplex virus thymidine kinase gene. EMBO J 1:1473–1477

    Google Scholar 

  • Reiss B, Sprengel R, Schaller H (1984a) Protein fusions with the kanamycin resistance gene from transposon Tn5. EMBO J 3:3317–3322

    Google Scholar 

  • Reiss B, Sprengel R, Will H, Schaller H (1984b) A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30:211–218

    Google Scholar 

  • Rochester DE, Winer JA, Shah DM (1986) The structure and expression of maize genes encoding the major heat shock protein, hsp70. EMBO J 5:451–458

    CAS  Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    CAS  PubMed  Google Scholar 

  • Schlesinger MJ, Ashburner M, Tissicres A (1982) Heat shock from bacteria to man. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Schöffl F, Raschke E, Nagao R (1984) The DNA sequence analysis of soybean heat-shock genes and identification of possible regulatory promoter elements. EMBO J 3:2491–2497

    Google Scholar 

  • Simon R, Priefer U, Pühler A (1983) A broad host range mobilization system for in vitro genetic engineering; transposon mutagenesis in Gram-negative bacteria. Bio/technology 1:784–791

    Google Scholar 

  • Sorger PK, Pelham HRB (1988) Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. Cell 54:855–864

    Google Scholar 

  • Spena A, Hain R, Ziervogel U, Saedler H, Schell J (1985) Construction of a heat-inducible gene for plants. Demonstration of heatinducible activity of the Drosophila hsp70 promoter in plants. EMBO J 4:2739–2743

    Google Scholar 

  • Storti RV, Scott MP, Rich A, Pardue ML (1980) Translational control of protein synthesis in response to heat shock in D. melanogaster cells. Cell 22:825–834

    Google Scholar 

  • Strittmatter G, Chua N-H (1987) Artifical combination of two cis-regulatory elements generates a unique pattern of expression in transgenic plants. Proc Natl Acad Sci USA 84:8986–8990

    Google Scholar 

  • Topol J, Ruden DM, Parker CS (1985) Sequences required for in vitro transcriptional activation of a Drosophila hsp70 gene. Cell 42:527–537

    Google Scholar 

  • Van den Elzen PJM, Townsend J, Lee KY, Bedbrook JR (1985) A chimaeric hygromycin resistance gene as a selectable marker in plant cells. Plant Mol Biol 5:299–302

    Google Scholar 

  • Velten J, Velten L, Hain R, Schell J (1984) Isolation of a dual plant promoter fragment from the Ti plasmid of Agrobacterium tumefaciens. EMBO J 3:2723–2730

    Google Scholar 

  • Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268

    Google Scholar 

  • Wiederrecht G, Shuey DJ, Kibbe WA, Parker CS (1987) The Saccharomyces and Drosophila heat shock transcription factors are identical in size and DNA binding properties. Cell 48:507–515

    Google Scholar 

  • Wiederrecht G, Seto D, Parker SS (1988) Isolation of the gene encoding the S. cerevisiae heat shock transcription factor. Cell 54:841–853

    Google Scholar 

  • Willmitzer L, Otten L, Simons G, Schmalenbach W, Schröder J, Schröder G, van Montagu M, de Vos G, Schell J (1981) Nuclear and polysomal transcripts of T-DNA in octopine crown gall suspension and callus cultures. Mol Gen Genet 182:255–262

    Google Scholar 

  • Wu C (1984) Two protein-binding sites in chromatin implicated in the activation of heat-shock genes. Nature 309:229–234

    Google Scholar 

  • Xiao H, Lis JT (1988) Germline transformation used to define key features of heat-shock response elements. Science 239:1139–1142

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 cloning vectors and hot strains: nucleotide sequences of the M13mp18 and pUC 19 vectors. Gene 33:103–119

    Article  CAS  PubMed  Google Scholar 

  • Zambryski P, Herrera-Estrella L, Block M, Van Montagu M, Schell J (1984) In: Hollaender A, Setlow J (eds) Genetic engineering, principles and methods, vol 6. Plenum, New York, pp 253–278

    Google Scholar 

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Communicated by H. Saedler

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Wing, D., Koncz, C. & Schell, J. Conserved function in Nicotiana tabacum of a single Drosophila hsp70 promoter heat shock element when fused to a minimal T-DNA promoter. Molec. Gen. Genet. 219, 9–16 (1989). https://doi.org/10.1007/BF00261151

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  • DOI: https://doi.org/10.1007/BF00261151

Key words

  • Heat shock
  • Plant gene transfer vector
  • Drosophila hsp70
  • Tobacco
  • Minimal promoter