Plant Molecular Biology

, Volume 56, Issue 1, pp 57–75 | Cite as

Plant class B HSFs inhibit transcription and exhibit affinity for TFIIB and TBP

  • Eva Czarnecka-verner
  • Songqin Pan
  • Tarek Salem
  • William B. Gurley


Plant heat shock transcription factors (HSFs) are capable of transcriptional activation (class A HSFs) or both, activation and repression (class B HSFs). However, the details of mechanism still remain unclear. It is likely, that the regulation occurs through interactions of HSFs with general transcription factors (GTFs), as has been described for numerous other transcription factors. Here, we show that class A HSFs may activate transcription through direct contacts with TATA-binding protein (TBP). Class A HSFs can also interact weakly with TFIIB. Conversely, class B HSFs inhibit promoter activity through an active mechanism of repression that involves the C-terminal regulatory region (CTR) of class B HSFs. Deletion analysis revealed two sites in the CTR of soybean GmHSFB1 potentially involved in protein–protein interactions with GTFs: one is the repressor domain (RD) located in the N-terminal half of the CTR, and the other is a TFIIB binding domain (BD) that shows affinity for TFIIB and is located C-terminally from the RD. A Gal4 DNA binding domain-RD fusion repressed activity of LexA-activators, while Gal4-BD proteins synergistically activated strong and weak transcriptional activators. In vitrobinding studies were consistent with this pattern of activity since the BD region alone interacted strongly with TFIIB, and the presence of RD had an inhibitory effect on TFIIB binding and transcriptional activation.

heat shock factor promoter protein interactions repression synergism 


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  1. Aranda, M.A., Escaler, M., Thomas, C.L., and Maule A.J. (1999). A heat stress transcription factor in pea is differentially controlled by heat and virus replication. Plant J. 20: 153–161.Google Scholar
  2. Baniahmad, A., Ha, I., Reinberg, D., Tsai, S., Tsai, M.-J. and O'Malley, B.W. (1993). Interaction of human hormone receptor beta with transcription factor TFIIB may mediate target gene derepression and activation by thyroid hormone. Proc. Natl. Acad. Sci. USA 90: 8832–8836.Google Scholar
  3. Berghagen, H., Ragnhildstveit, E., Krogsrud, K., Thuestad, G., Apriletti, J. and Saatcioglu, F. (2002). Corepressor SMRT functions as a coactivator for thyroid hormone receptor T3Ralpha from a negative hormone response element. J. Biol. Chem. 277: 49517–49522.Google Scholar
  4. Bharti, K., von Koskull-Doring, P., Bharti, S., Kumar, P., Tintschl-Korbitzer, A., Treuter, E. and Nover, L. (2004). Tomato heat stress transcription factor HsfB1 represents a novel type of general transcriptional coactivator with a histone-like motif interacting with the plant CREB binding protein ortholog HAC1. Plant Cell 16: 1521–1535.Google Scholar
  5. Corton, J.C., Moreno, E. and Johnston, S.A. (1998). Alterations in the GAL4 DNA-binding domain can affect transcriptional activation independent of DNA binding. J. Biol. Chem. 273: 13776–13780.Google Scholar
  6. Czarnecka-Verner, E. and Gurley, W.B. (1999). Plant heat shock transcription factors: divergence in structure and function. Biotechnologia 3: 125–142.Google Scholar
  7. Czarnecka-Verner, E. and Gurley, W.B. (2002). Arabidopsis class A and B HSFs show a spectrum of transcriptional activity. Biotechnologia 3: 15–27.Google Scholar
  8. Czarnecka-Verner, E., Pan, S., Yuan, C.-X. and Gurley, W.B. (2000). Functional specialization of plant class A and B HSFs. In: J.H. Cherry (Ed.), Plant Tolerance to Abiotic Stresses in Agriculture: Role of Genetic Engineering. Kluwer Academic Publishers, The Netherlands, pp. 3–28.Google Scholar
  9. Czarnecka-Verner, E., Yuan, C.X., Fox, P.C. and Gurley, W.B. (1995). Isolation and characterization of six heat shock transcription factor genes from soybean. Plant Mol. Biol. 29: 37–51.Google Scholar
  10. Czarnecka-Verner, E., Yuan C.-X., Nover, L., Scharf, K.-D., Englich, G. and Gurley, W.B. (1998). Plant heat shock transcription factors: positive and negative aspects of regulation. Acta Physiol. Plant. 19: 529–537.Google Scholar
  11. Czarnecka-Verner, E., Yuan, C.-X., Scharf, K.-D., Englich, G. and Gurley, W.B. (2000). Plants contain a novel multimember class of heat shock factors without transcriptional activation potential. Plant Mol. Biol. 43: 459–471.Google Scholar
  12. Eckey, M., Moehren, U. and Baniahmad, A. (2003). Gene silencing by the thyroid hormone receptor. Mol. Cell. Endocrinol. 213: 13–22Google Scholar
  13. Fondell, J.D., Brunel, F., Hisatake, K. and Roeder, R.G. (1995). Unliganded thyroid hormone receptor alpha can target TATA-binding protein for transcriptional repression. Mol. Cell. Biol. 16: 281–287.Google Scholar
  14. Fondell, J.D., Roy A.L. and Roeder, R.G. (1993). Unliganded thyroid hormone receptor inhibits formation of a functional preinitiation complex: implications for active repression. Genes Dev. 7: 1400–1410.Google Scholar
  15. Frejtag, W., Zhang, Y., Dai, R., Anderson, M.G. and Mivechi, N.F. (2001). Heat shock factor 4 (HSF-4a) represses basal transcription through interaction with TFIIF. J. Biol. Chem. 276: 14685–14694.Google Scholar
  16. Gagliardi, D., Breton, C., Chaboud, A., Vergne, P. and Dumas, C. (1995). Expression of heat shock factor and heat shock protein 70 genes during maize pollen development. Plant Mol. Biol. 29: 841–856.Google Scholar
  17. Hall, D.B. and Struhl, K. (2002). The VP16 activation domain interacts with multiple transcriptional components as determined by protein-protein cross-linking in vivo. J. Biol. Chem. 277: 46043–46050.Google Scholar
  18. Hanna-Rose, W. and Hansen, U. (1996) Active repression mechanisms of eukaryotic transcription repressors. Trends Genet. 12: 229–234.Google Scholar
  19. Higgins, D., Thompson, J., Gibson, T., Thompson, J.D., Higgins, D.G. and Gibson, T.J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673–4680.Google Scholar
  20. Hübel, A. and Schöffl, F. (1994). Arabidopsis heat shock factor: isolation and characterization of the gene and the recombinant protein. Plant Mol. Biol. 26: 353–362.Google Scholar
  21. Key, J.L., Kimpel, J., Vierling, E., Lin, C.-Y., Nagao, R.T., Czarnecka, E. and Schoffl, F. (1985). Physiological and molecular analyses of the heat shock response in plants. In: B.G. Atkinson and D.B. Walden (Eds.) Changes in Eukaryotic Gene Expression in Response to Environmental Stress. Academic Press, Inc. pp. 327–348.Google Scholar
  22. Kuras, L., Kosa, P., Mencia, M. and Struhl, K. (2000). TAFcontaining and TAF-independent forms of transcriptionally active TBP in vivo. Science 288: 1244–1248.Google Scholar
  23. Lin, Y.-S. and Green, M.R. (1991). Mechanism of action of an acidic transcriptional activator in vitro. Cell 64: 971–981.Google Scholar
  24. Lyck, R., Harmening, U., Hohfeld, I., Treuter, E., Scharf, K.-D. and Nover, L. (1997). Intracellular distribution and identification of the nuclear localization signals of two plant heat-stress transcription factors. Planta 202: 117–125.Google Scholar
  25. Mason, P.B., Jr. and Lis, J.T. (1997). Cooperative and competitive protein interactions at the hsp70 promoter. J. Biol. Chem. 272: 33227–33233.Google Scholar
  26. Mishra, A.K., Vasanthi, P. and Bhargava, P. (2003). The transcriptional activator Gal4-VP16 regulates the intramolecular interactions of the TATA-binding protein. J. Biosci. 28: 423–426.Google Scholar
  27. Muscat, G.E.O., Burke, L.J. and Downes, M. (1998). The corepressor N-CoR and its variants RIP13a and RIP13D1 directly interact with the basal transcription factors TFIIB, TAFII32 and TAFII70. Nucleic Acids Res. 26: 2899–2907.Google Scholar
  28. Nedialkov, Y.A. and Triezenberg, S.J. (2004). Quantitative assessment of in vitro interactions implicates TATA-binding protein as a target of the VP16C transcriptional activation region. Arch. Biochem. Biophys. 425: 77–86.Google Scholar
  29. Nover, L., Bharti, K., Doring, P., Mishra, S.K., Ganguli, A. and Scharf, K.-D. (2001). Arabidopsis and the heat stress transcription factor world: how many heat stress transcription factors do we need? Cell Stress & Chap 6: 177–189.Google Scholar
  30. Nover, L., Scharf, K.-D., Gagliardi, D., Vergne, P., Czarnecka-Verner, E. and Gurley, W.B. (1996). The Hsf world: classification and properties of plant heat stress transcription factors. Cell Stress & Chap 1: 215–223.Google Scholar
  31. Pan, S., Sehnke, P.C., Ferl, R.J. and Gurley, W.B. (1999). Specific interactions with TBP and TFIIB in vitro suggest that 14–3–3 proteins may participate in the regulation of transcription when part of a DNA binding complex. Plant Cell 11: 1591–1602.Google Scholar
  32. Rasmussen, E.B. and Lis, J.T. (1993). In vivo transcriptional pausing and cap formation on three Drosophila heat shock genes. Proc. Natl. Acad. Sci. USA 90: 7923–7927.Google Scholar
  33. Reindl, A. and Schöffl, F. (1998). Interaction between the Arabidopsis thaliana heat shock transcription factor HSF1 and the TATA binding protein TBP. FEBS Lett. 436: 318–322.Google Scholar
  34. Roberts, S.G.E. (2000). Mechanisms of action of transcriptional activation and repression domains. Cell. Mol. Life Sci. 57: 1149–1160.Google Scholar
  35. Scharf, K.D., Heider, H., Hohfeld, I., Lyck, R., Schmidt, E. and Nover, L. (1998). The tomato Hsf system: HsfA2 needs interaction with HsfA1 for efficient nuclear import and may be localized in cytoplasmic heat stress granules. Mol. Cell. Biol. 18: 2240–2251.Google Scholar
  36. Scharf, K.D., Rose, S., Zott, W., Schoffl, F. and Nover, L. (1990). Three tomato genes code for heat stress transcription factors with a region of remarkable homology to the DNAbinding domain of the yeast HSF [published erratum appears in EMBO J 1991: 1026]. EMBO J 9: 4495–4501.Google Scholar
  37. Shoji, T., Kato, K., Sekine, M., Yosida, K. and Shinmyo, A. (2000). Two types of heat stress factors in cultured tobacco cells. Plant Cell Rep. 19: 414–420.Google Scholar
  38. Tong, G.X., Tanen, M.R. and Bagchi, M.K. (1995). Ligand modulates the interaction of thyroid hormone receptor beta with the basal transcription machinery. J. Biol. Chem. 270: 10601–10611.Google Scholar
  39. Treuter, E., Nover, L., Ohme, K. and Scharf, K.-D. (1993). Promoter specificity and deletion analysis of three heat stress transcription factors of tomato. Mol. Gen. Genet. 240: 113–125.Google Scholar
  40. Usheva, A., Maldonado, E., Goldring, A., Lu, H., Houbavi, C., Reinberg, D. and Aloni, Y. (1992). Specific interaction between the nonphosphorylated form of RNA polymerase II and the TATA-binding protein. Cell 69: 871–881.Google Scholar
  41. Xiao, H., Friesen, J.D. and Lis, J.L. (1994). A highly conserved domain of RNA polymerase II shares a functional element with acidic activation domains of upstream transcription factors. Mol. Cell. Biol. 14: 7507–7516.Google Scholar
  42. Yuan, C.-X. and Gurley, W.B. (2000). Potential targets for HSF1 within the preinitiation complex. Cell Stress & Chap 5: 229–242.Google Scholar
  43. Zhu, Q., Ordiz, M.I., Dabi, T., Beachy, R.N. and Lamb, C. (2002). Rice TATA binding protein interacts functionally with transcription factor IIB and the RF2a bZIP transcriptional activator in an enhanced plant in vitro transcription system. Plant Cell 14: 795–803.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Eva Czarnecka-verner
    • 1
  • Songqin Pan
    • 1
  • Tarek Salem
    • 1
  • William B. Gurley
    • 1
  1. 1.Microbiology and Cell Science Department, Program of Plant Molecular and Cellular BiologyUniversity of FloridaGainesvilleUSA

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