Plant Molecular Biology

, Volume 29, Issue 1, pp 37–51

Isolation and characterization of six heat shock transcription factor cDNA clones from soybean

  • Eva Czarnecka-Verner
  • Chao-Xing Yuan
  • Paul C. Fox
  • William B. Gurley
Research Article


Thermal stress in soybean seedlings causes the activation of pre-existing heat shock transcription factor proteins (HSFs). Activation results in the induction of DNA binding activity which leads to the transcription of heat shock genes. From a soybean cDNA library we have isolated cDNA clones corresponding to six HSF genes. Two HSF genes are expressed constitutively at the transcriptional level, and the remaining four are heat-inducible. Two of the heat inducible genes are also responsive to cadmium stress. Comparative analysis of HSF sequences indicated higher conservation of the DNA binding domain among plant HSFs than those from yeast or other higher eukaryotes. The putative plant HSF oligomerization domain contains hydrophobic heptapeptide repeats characteristic of coiled coils and seems to exist in two structural variants. The carboxy-terminal domains are reduced in size and the C-terminal heptad repeat is degenerate.

Key words

HSF DNA binding domain oligomerization domain stress 


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  1. 1.
    AlberT: Structure of the leucine zipper. Curr Opin Genet Devel 2: 205–210 (1992).Google Scholar
  2. 2.
    ChenY, BarlevNA, WestergaardO, JakobsenBK: Identification of the C-terminal activator domain in yeast heat shock factor: independent control of transient and sustained transcriptional activity. EMBO J 12: 5007–5018 (1993).PubMedGoogle Scholar
  3. 3.
    ClosJ, WestwoodJT, BeckerPB, WilsonS, LambertK, WuC: Molecular cloning and expression of hexameric Drosophila heat shock factor subject to negative regulation. Cell 63: 1085–1097 (1990).CrossRefPubMedGoogle Scholar
  4. 4.
    CzarneckaE, EdelmanL, SchöfflF, KeyJL: Comparative analysis of physical stress responses in soybean seedlings using cloned heat shock cDNAs (Glycine max). Plant Mol Biol 3: 45–58 (1984).Google Scholar
  5. 5.
    CzarneckaE, FoxPC, GurleyWB: In vitro interaction of nuclear proteins with the promoter of soybean heat shock gene Gmhsp17.5E. Plant Physiol 94: 935–943 (1990).Google Scholar
  6. 6.
    CzarneckaE, IngersollJC, GurleyWB: AT-rich promoter elements of soybean heat shock gene Gmhsp17.5E bind two distinct sets of proteins in vitro. Plant Mol Biol 19: 985–1000 (1992).PubMedGoogle Scholar
  7. 7.
    CzarneckaE, NagaoRT, KeyJL, GurleyWB: Characterization of Gmhsp26-A, a stress gene encoding a divergent heat shock protein from soybean: heavy-metal-induced inhibition of intron processing. Mol Cell Biol 8: 1113–1122 (1988).PubMedGoogle Scholar
  8. 8.
    Czarnecka-VernerE, BarrosMD, GurleyWB: Regulation of heat shock gene expression. In: BasraAS (eds) Stress-Induced Gene Expression in Plants, pp. 131–161. Hardwood Academic Publishers, Switzerland (1994).Google Scholar
  9. 9.
    FernandesM, O'BrianT, LisJT: Structure and regulation of heat shock gene promoters. In: MorimotoRI, TissièresA, GeorgopoulosC (eds) The Biology of Heat Shock Proteins and Molecular Chaperones, pp. 375–393. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1994).Google Scholar
  10. 10.
    FlickKE, GonzalezAJ, HarrisonCJ, NelsonHCM: Yeast heat shock transcription factor contains a flexible linker between the DNA-binding and trimerization domains. J Biol Chem 269: 12475–12481 (1994).PubMedGoogle Scholar
  11. 11.
    FrankelS, SohnR, LeinwandL: The use of sarcosyl in generating soluble protein after bacterial expression. Proc Natl Acad Sci USA 88: 1192–1196 (1991).PubMedGoogle Scholar
  12. 12.
    HarrisonCJ, BohmAA, NelsonHCM: Crystal structure of the DNA binding domain of the heat shock transcription factor. Science 263: 224–227 (1994).PubMedGoogle Scholar
  13. 13.
    HuJC, SauerRT: The basic-region leucine-zipper family of DNA binding proteins. In: EcksteinF, LilleyDMJ (eds) Nucleic Acids and Molecular Biology, vol. 6, pp. 82–101. Springer-Verlag, Berlin/Heidelberg (1992).Google Scholar
  14. 14.
    HübelA, SchöfflF: Arabidopsis heat shock factor: isolation and characterization of the gene and the recombinant protein. Plant Mol Biol 26: 353–362 (1994).PubMedGoogle Scholar
  15. 15.
    JakobsenBK, PelhamHRB: A conserved heptapeptide restrains the activity of the yeast heat shock transcription factor. EMBO J 10: 369–375 (1991).PubMedGoogle Scholar
  16. 16.
    JofukuKD, GoldbergRB: Analysis of plant gene structure. In: ShawCH (eds) Plant Molecular Biology: A Practical Approach, pp. 37–66. IRL Press, Eynsham, Oxford, England (1988).Google Scholar
  17. 17.
    LaemmliUK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).PubMedGoogle Scholar
  18. 18.
    MorimotoRI, JurivichDA, KroegerPE, MathurSK, MurphySP, NakaiA, SargeK, AbravayaK, SistonenLT: Regulation of heat shock gene transcription by a family of heat shock factors. In: MorimotoRI, TissièresA, GeorgopoulosC (eds) The Biology of Heat Shock Proteins and Molecular Chaperones, pp. 417–455. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1994).Google Scholar
  19. 19.
    NakaiA, MorimotoRI: Characterization of a novel chicken heat shock transcription factor, heat shock factor 3, suggests a new regulatory pathway. Mol Cell Biol 13: 1983–1997 (1993).PubMedGoogle Scholar
  20. 20.
    PeteranderlR, NelsonHCM: Trimerization of the heat shock transcription factor by a triple-stranded α-helical coiled coil. Biochemistry 31: 12272–12276 (1992).PubMedGoogle Scholar
  21. 21.
    RabindranSK, GiorgiG, ClosJ, WuC: Molecular cloning and expression of a human heat shock factor, HSF1. Proc Natl Acad Sci USA 88: 6906–6910 (1991).PubMedGoogle Scholar
  22. 22.
    RabindranSK, HarounRI, ClosJ, WisniewskiJ, WuC: Regulation of heat shock factor trimer formation: role of a conserved leucine zipper. Science 259: 230–234 (1993).PubMedGoogle Scholar
  23. 23.
    SangerF, NicklenS, CoulsonAR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).PubMedGoogle Scholar
  24. 24.
    SargeKD, ZimarinoV, HolmK, WuC, MorimotoRJ: Cloning and characterization of two mouse heat shock factors with inducible and constitutive DNA-binding ability. Genes Devel 5: 1902–1911 (1991).Google Scholar
  25. 25.
    ScharfK-D, MaternaT, TreuterE, NoverL: Heat stress promoters and transcription factors. In: NoverL (eds) Plant Promoters and Transcription Factors, pp. 121–158. Springer-Verlag, Berlin/Heidelberg (1994).Google Scholar
  26. 26.
    ScharfK-D, RoseS, ThierfelderJ, NoverL: Two cDNAs for tomato heat stress transcription factors. Plant Physiol 102: 1355–1356 (1993).CrossRefPubMedGoogle Scholar
  27. 27.
    ScharfK-D, RoseS, ZottW, SchöfflF, NoverL: Three tomato genes code for heat stress transcription factors with a region of remarkable homology to the DNA-binding domain of the yeast HSF. EMBO J 9: 4495–4501 (1990).PubMedGoogle Scholar
  28. 28.
    SchuetzTJ, GalloGJ, SheldonL, TempstP, KingstonRE: Isolation of a cDNA for HSF2: evidence for two heat shock factor genes in humans. Proc Natl Acad Sci USA 88: 6910–6915 (1991).Google Scholar
  29. 29.
    SheldonLA, KingstonRE: Hydrophobic coiled-coil domains regulate the subcellular localization of human heat shock factor 2. Genes Devel 7: 1549–1558 (1993).PubMedGoogle Scholar
  30. 30.
    SistonenL, SargeKD, PhillipsB, AbravayaK, MorimotoRI: Activation of heat shock factor 2 during hemin-induced differentiation of human erythroleukemia cells. Mol Cell Biol 12: 4104–4111 (1992).Google Scholar
  31. 31.
    TreuterE, NoverL, OhmeK, ScharfK-D: Promoter specificity and deletion analysis of three heat stress transcription factors of tomato. Mol Gen Genet 240: 113–125 (1993).Google Scholar
  32. 32.
    VuisterGW, KimS, WuC, BaxA: NMR evidence for similarities between the DNA-binding regions of Drosophila melanogaster heat shock factor and the helix-turn-helix and HNF-3/forkhead families of transcription factors. Biochemistry 33: 10–16 (1994).PubMedGoogle Scholar
  33. 33.
    WiederrechtG, SietoD, ParkerCS: Isolation of the gene encoding the S. cerevisiae heat shock transcription factor. Cell 54: 841–853 (1988).CrossRefPubMedGoogle Scholar
  34. 34.
    WuC, ClosJ, GiorgiG, HarounRI, RimS-J, RabindranSK, WestwoodJT, WisniewskiJ, YimG: Structure and regulation of heat shock transcription factor. In: MorimotoRI, TissièresA, GeorgopoulosC (eds) The Biology of Heat Shock Proteins and Molecular Chaperones, pp. 395–416. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1994).Google Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Eva Czarnecka-Verner
    • 1
  • Chao-Xing Yuan
    • 1
  • Paul C. Fox
    • 1
  • William B. Gurley
    • 1
  1. 1.Department of Microbiology and Cell Science, Program of Plant Molecular and Cellular BiologyUniversity of FloridaGainesvilleUSA

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