Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1996 Mar;16(3):839–846. doi: 10.1128/mcb.16.3.839

The regulatory domain of human heat shock factor 1 is sufficient to sense heat stress.

E M Newton 1, U Knauf 1, M Green 1, R E Kingston 1
PMCID: PMC231064  PMID: 8622685

Abstract

Heat shock factor (HSF) activates transcription in response to cellular stress. Human HSF1 has a central regulatory domain which can repress the activity of its activation domains at the control temperature and render them heat shock inducible. To determine whether the regulatory domain works in tandem with specific features of the HSF1 transcriptional activation domains, we first used deletion and point mutagenesis to define these activation domains. One of the activation domains can be reduced to just 20 amino acids. A GAL4 fusion protein containing the HSF 1 regulatory domain and this 20-amino-acid activation domain is repressed at the control temperature but potently activates transcription in response to heat shock. No specific amino acids in this activation domain are required for response to the regulatory domain; in particular, none of the potentially phosphorylated serine and threonine residues are required for heat induction, implying that heat-induced phosphorylation of the transcriptional activation domains is not required for induction. The regulatory domain is able to confer heat responsiveness to an otherwise completely heterologous chimeric activator that contains a portion of the VP16 activation domain, suggesting that the regulatory domain can sense heat in the absence of other portions of HSF1.

Full Text

The Full Text of this article is available as a PDF (529.9 KB).

Selected References

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

  1. Bonner J. J., Heyward S., Fackenthal D. L. Temperature-dependent regulation of a heterologous transcriptional activation domain fused to yeast heat shock transcription factor. Mol Cell Biol. 1992 Mar;12(3):1021–1030. doi: 10.1128/mcb.12.3.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bruce J. L., Price B. D., Coleman C. N., Calderwood S. K. Oxidative injury rapidly activates the heat shock transcription factor but fails to increase levels of heat shock proteins. Cancer Res. 1993 Jan 1;53(1):12–15. [PubMed] [Google Scholar]
  3. Cress W. D., Triezenberg S. J. Critical structural elements of the VP16 transcriptional activation domain. Science. 1991 Jan 4;251(4989):87–90. doi: 10.1126/science.1846049. [DOI] [PubMed] [Google Scholar]
  4. Gallo G. J., Schuetz T. J., Kingston R. E. Regulation of heat shock factor in Schizosaccharomyces pombe more closely resembles regulation in mammals than in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Jan;11(1):281–288. doi: 10.1128/mcb.11.1.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Giardina C., Lis J. T. Dynamic protein-DNA architecture of a yeast heat shock promoter. Mol Cell Biol. 1995 May;15(5):2737–2744. doi: 10.1128/mcb.15.5.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Green M., Schuetz T. J., Sullivan E. K., Kingston R. E. A heat shock-responsive domain of human HSF1 that regulates transcription activation domain function. Mol Cell Biol. 1995 Jun;15(6):3354–3362. doi: 10.1128/mcb.15.6.3354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hensold J. O., Hunt C. R., Calderwood S. K., Housman D. E., Kingston R. E. DNA binding of heat shock factor to the heat shock element is insufficient for transcriptional activation in murine erythroleukemia cells. Mol Cell Biol. 1990 Apr;10(4):1600–1608. doi: 10.1128/mcb.10.4.1600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hightower L. E. Heat shock, stress proteins, chaperones, and proteotoxicity. Cell. 1991 Jul 26;66(2):191–197. doi: 10.1016/0092-8674(91)90611-2. [DOI] [PubMed] [Google Scholar]
  9. Hope I. A., Struhl K. GCN4, a eukaryotic transcriptional activator protein, binds as a dimer to target DNA. EMBO J. 1987 Sep;6(9):2781–2784. doi: 10.1002/j.1460-2075.1987.tb02573.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Høj A., Jakobsen B. K. A short element required for turning off heat shock transcription factor: evidence that phosphorylation enhances deactivation. EMBO J. 1994 Jun 1;13(11):2617–2624. doi: 10.1002/j.1460-2075.1994.tb06552.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jakobsen B. K., Pelham H. R. A conserved heptapeptide restrains the activity of the yeast heat shock transcription factor. EMBO J. 1991 Feb;10(2):369–375. doi: 10.1002/j.1460-2075.1991.tb07958.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jakobsen B. K., Pelham H. R. Constitutive binding of yeast heat shock factor to DNA in vivo. Mol Cell Biol. 1988 Nov;8(11):5040–5042. doi: 10.1128/mcb.8.11.5040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jurivich D. A., Sistonen L., Kroes R. A., Morimoto R. I. Effect of sodium salicylate on the human heat shock response. Science. 1992 Mar 6;255(5049):1243–1245. doi: 10.1126/science.1546322. [DOI] [PubMed] [Google Scholar]
  14. Jurivich D. A., Sistonen L., Sarge K. D., Morimoto R. I. Arachidonate is a potent modulator of human heat shock gene transcription. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2280–2284. doi: 10.1073/pnas.91.6.2280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kingston R. E., Cowie A., Morimoto R. I., Gwinn K. A. Binding of polyomavirus large T antigen to the human hsp70 promoter is not required for trans activation. Mol Cell Biol. 1986 Sep;6(9):3180–3190. doi: 10.1128/mcb.6.9.3180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kingston R. E., Schuetz T. J., Larin Z. Heat-inducible human factor that binds to a human hsp70 promoter. Mol Cell Biol. 1987 Apr;7(4):1530–1534. doi: 10.1128/mcb.7.4.1530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Larson J. S., Schuetz T. J., Kingston R. E. Activation in vitro of sequence-specific DNA binding by a human regulatory factor. Nature. 1988 Sep 22;335(6188):372–375. doi: 10.1038/335372a0. [DOI] [PubMed] [Google Scholar]
  18. Larson J. S., Schuetz T. J., Kingston R. E. In vitro activation of purified human heat shock factor by heat. Biochemistry. 1995 Feb 14;34(6):1902–1911. doi: 10.1021/bi00006a011. [DOI] [PubMed] [Google Scholar]
  19. Lee K. A., Bindereif A., Green M. R. A small-scale procedure for preparation of nuclear extracts that support efficient transcription and pre-mRNA splicing. Gene Anal Tech. 1988 Mar-Apr;5(2):22–31. doi: 10.1016/0735-0651(88)90023-4. [DOI] [PubMed] [Google Scholar]
  20. Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151–1191. doi: 10.1146/annurev.bi.55.070186.005443. [DOI] [PubMed] [Google Scholar]
  21. Picard D., Salser S. J., Yamamoto K. R. A movable and regulable inactivation function within the steroid binding domain of the glucocorticoid receptor. Cell. 1988 Sep 23;54(7):1073–1080. doi: 10.1016/0092-8674(88)90122-5. [DOI] [PubMed] [Google Scholar]
  22. Price B. D., Calderwood S. K. Ca2+ is essential for multistep activation of the heat shock factor in permeabilized cells. Mol Cell Biol. 1991 Jun;11(6):3365–3368. doi: 10.1128/mcb.11.6.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Regier J. L., Shen F., Triezenberg S. J. Pattern of aromatic and hydrophobic amino acids critical for one of two subdomains of the VP16 transcriptional activator. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):883–887. doi: 10.1073/pnas.90.3.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sarge K. D., Murphy S. P., Morimoto R. I. Activation of heat shock gene transcription by heat shock factor 1 involves oligomerization, acquisition of DNA-binding activity, and nuclear localization and can occur in the absence of stress. Mol Cell Biol. 1993 Mar;13(3):1392–1407. doi: 10.1128/mcb.13.3.1392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sheldon L. A., Kingston R. E. Hydrophobic coiled-coil domains regulate the subcellular localization of human heat shock factor 2. Genes Dev. 1993 Aug;7(8):1549–1558. doi: 10.1101/gad.7.8.1549. [DOI] [PubMed] [Google Scholar]
  26. Shi Y., Kroeger P. E., Morimoto R. I. The carboxyl-terminal transactivation domain of heat shock factor 1 is negatively regulated and stress responsive. Mol Cell Biol. 1995 Aug;15(8):4309–4318. doi: 10.1128/mcb.15.8.4309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sorger P. K., Lewis M. J., Pelham H. R. Heat shock factor is regulated differently in yeast and HeLa cells. Nature. 1987 Sep 3;329(6134):81–84. doi: 10.1038/329081a0. [DOI] [PubMed] [Google Scholar]
  28. Sorger P. K., Pelham H. R. Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. Cell. 1988 Sep 9;54(6):855–864. doi: 10.1016/s0092-8674(88)91219-6. [DOI] [PubMed] [Google Scholar]
  29. Sorger P. K. Yeast heat shock factor contains separable transient and sustained response transcriptional activators. Cell. 1990 Aug 24;62(4):793–805. doi: 10.1016/0092-8674(90)90123-v. [DOI] [PubMed] [Google Scholar]
  30. Wu C. Activating protein factor binds in vitro to upstream control sequences in heat shock gene chromatin. Nature. 1984 Sep 6;311(5981):81–84. doi: 10.1038/311081a0. [DOI] [PubMed] [Google Scholar]
  31. Zimarino V., Tsai C., Wu C. Complex modes of heat shock factor activation. Mol Cell Biol. 1990 Feb;10(2):752–759. doi: 10.1128/mcb.10.2.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zimarino V., Wu C. Induction of sequence-specific binding of Drosophila heat shock activator protein without protein synthesis. 1987 Jun 25-Jul 1Nature. 327(6124):727–730. doi: 10.1038/327727a0. [DOI] [PubMed] [Google Scholar]
  33. Zuo J., Rungger D., Voellmy R. Multiple layers of regulation of human heat shock transcription factor 1. Mol Cell Biol. 1995 Aug;15(8):4319–4330. doi: 10.1128/mcb.15.8.4319. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES