Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1986 Jul;5(7):1653–1658. doi: 10.1002/j.1460-2075.1986.tb04408.x

Activation of the Drosophila hsp27 promoter by heat shock and by ecdysone involves independent and remote regulatory sequences

Guy Riddihough 1, Hugh RB Pelham 1
PMCID: PMC1166991  PMID: 16453691

Abstract

Transcription of the Drosophila hsp27 gene is induced both by heat shock and by the steroid hormone ecdysone. Deletion analysis of the promoter using a transient expression assay in Drosophila tissue culture cells reveals two separate and functionally independent regulatory regions, whose positions coincide with previously reported DNase I-hypersensitive sites in embryonic chromatin. Ecdysone induction is mediated by multiple elements between −579 and −455. Several matches to the consensus heat shock regulatory element (HSE) lie between −370 and −270, and deletion of these abolishes the response to heat shock. Heat inducibility can be restored by the addition of HSE sequences from the hsp70 promoter, even when these are placed at the 3' end of the gene. Furthermore, HSEs seperated by >2 kb can interact in a cooperative way to promote transcription. Deletion of sequences adjacent to the TATA box (−40 to −154) has little effect on induction by either heat shock or ecdysone. In this respect, activation of the hsp27 TATA box in Drosophila cells differs from the long-range activation of promoters by HSEs or viral enhancers in HeLa cells.

Keywords: ecdysone, Drosophila, heat shock, transcription

Full text

PDF
1653

Images in this article

1654Fig. 1.
on p.1654
1654Fig. 3.
on p.1654
1656Fig. 8.
on p.1656

Selected References

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

  1. Ayme A., Southgate R., Tissières A. Nucleotide sequences responsible for the thermal inducibility of the Drosophila small heat-shock protein genes in monkey COS cells. J Mol Biol. 1985 Apr 20;182(4):469–475. doi: 10.1016/0022-2836(85)90233-5. [DOI] [PubMed] [Google Scholar]
  2. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Biggin M., Farrell P. J., Barrell B. G. Transcription and DNA sequence of the BamHI L fragment of B95-8 Epstein-Barr virus. EMBO J. 1984 May;3(5):1083–1090. doi: 10.1002/j.1460-2075.1984.tb01933.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  5. Cohen R. S., Meselson M. Separate regulatory elements for the heat-inducible and ovarian expression of the Drosophila hsp26 gene. Cell. 1985 Dec;43(3 Pt 2):737–746. doi: 10.1016/0092-8674(85)90247-8. [DOI] [PubMed] [Google Scholar]
  6. Costlow N., Lis J. T. High-resolution mapping of DNase I-hypersensitive sites of Drosophila heat shock genes in Drosophila melanogaster and Saccharomyces cerevisiae. Mol Cell Biol. 1984 Sep;4(9):1853–1863. doi: 10.1128/mcb.4.9.1853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dierks P., van Ooyen A., Cochran M. D., Dobkin C., Reiser J., Weissmann C. Three regions upstream from the cap site are required for efficient and accurate transcription of the rabbit beta-globin gene in mouse 3T6 cells. Cell. 1983 Mar;32(3):695–706. doi: 10.1016/0092-8674(83)90055-7. [DOI] [PubMed] [Google Scholar]
  8. Dudler R., Travers A. A. Upstream elements necessary for optimal function of the hsp 70 promoter in transformed flies. Cell. 1984 Sep;38(2):391–398. doi: 10.1016/0092-8674(84)90494-x. [DOI] [PubMed] [Google Scholar]
  9. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hall C. V., Jacob P. E., Ringold G. M., Lee F. Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells. J Mol Appl Genet. 1983;2(1):101–109. [PubMed] [Google Scholar]
  11. Hochschild A., Ptashne M. Cooperative binding of lambda repressors to sites separated by integral turns of the DNA helix. Cell. 1986 Mar 14;44(5):681–687. doi: 10.1016/0092-8674(86)90833-0. [DOI] [PubMed] [Google Scholar]
  12. Hoffman E., Corces V. Sequences involved in temperature and ecdysterone-induced transcription are located in separate regions of a Drosophila melanogaster heat shock gene. Mol Cell Biol. 1986 Feb;6(2):663–673. doi: 10.1128/mcb.6.2.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ingolia T. D., Craig E. A. Primary sequence of the 5' flanking regions of the Drosophila heat shock genes in chromosome subdivision 67B. Nucleic Acids Res. 1981 Apr 10;9(7):1627–1642. doi: 10.1093/nar/9.7.1627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ireland R. C., Berger E., Sirotkin K., Yund M. A., Osterbur D., Fristrom J. Ecdysterone induces the transcription of four heat-shock genes in Drosophila S3 cells and imaginal discs. Dev Biol. 1982 Oct;93(2):498–507. doi: 10.1016/0012-1606(82)90137-3. [DOI] [PubMed] [Google Scholar]
  15. Lawson R., Mestril R., Luo Y., Voellmy R. Ecdysterone selectively stimulates the expression of a 23000-Da heat-shock protein-beta-galactosidase hybrid gene in cultured Drosophila cells. Dev Biol. 1985 Aug;110(2):321–330. doi: 10.1016/0012-1606(85)90091-0. [DOI] [PubMed] [Google Scholar]
  16. Morganelli C. M., Berger E. M., Pelham H. R. Transcription of Drosophila small hsp-tk hybrid genes is induced by heat shock and by ecdysterone in transfected Drosophila cells. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5865–5869. doi: 10.1073/pnas.82.17.5865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nagawa F., Fink G. R. The relationship between the "TATA" sequence and transcription initiation sites at the HIS4 gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8557–8561. doi: 10.1073/pnas.82.24.8557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Parker C. S., Topol J. A Drosophila RNA polymerase II transcription factor binds to the regulatory site of an hsp 70 gene. Cell. 1984 May;37(1):273–283. doi: 10.1016/0092-8674(84)90323-4. [DOI] [PubMed] [Google Scholar]
  19. Pelham H. R. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene. Cell. 1982 Sep;30(2):517–528. doi: 10.1016/0092-8674(82)90249-5. [DOI] [PubMed] [Google Scholar]
  20. Pelham H. R., Bienz M. A synthetic heat-shock promoter element confers heat-inducibility on the herpes simplex virus thymidine kinase gene. EMBO J. 1982;1(11):1473–1477. doi: 10.1002/j.1460-2075.1982.tb01340.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Struhl K. The yeast his3 promoter contains at least two distinct elements. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7385–7389. doi: 10.1073/pnas.79.23.7385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Topol J., Ruden D. M., Parker C. S. Sequences required for in vitro transcriptional activation of a Drosophila hsp 70 gene. Cell. 1985 Sep;42(2):527–537. doi: 10.1016/0092-8674(85)90110-2. [DOI] [PubMed] [Google Scholar]
  23. Vitek M. P., Berger E. M. Steroid and high-temperature induction of the small heat-shock protein genes in Drosophila. J Mol Biol. 1984 Sep 15;178(2):173–189. doi: 10.1016/0022-2836(84)90138-4. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Wu C. An exonuclease protection assay reveals heat-shock element and TATA box DNA-binding proteins in crude nuclear extracts. Nature. 1985 Sep 5;317(6032):84–87. doi: 10.1038/317084a0. [DOI] [PubMed] [Google Scholar]
  26. Wu C. Two protein-binding sites in chromatin implicated in the activation of heat-shock genes. Nature. 1984 May 17;309(5965):229–234. doi: 10.1038/309229a0. [DOI] [PubMed] [Google Scholar]
  27. Zimmerman J. L., Petri W., Meselson M. Accumulation of a specific subset of D. melanogaster heat shock mRNAs in normal development without heat shock. Cell. 1983 Apr;32(4):1161–1170. doi: 10.1016/0092-8674(83)90299-4. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES