Abstract
The 5' DNA sequences involved in the thermal inducibility of the soybean heat shock gene hs6871 were analysed in transgenic tobacco plants. The transcriptional activity of various in vitro generated deletion mutants was examined by Northern blot analysis, S1 nuclease mapping and dot-blot hybridization. At least 181 bp upstream from the translational start site are sufficient for thermal induction at 40°C and correct initiation of transcripts. Full promoter activity with the induction of wild-type levels of transcripts requires additional upstream sequences contained within 439 bp 5' to the coding sequence. Our results suggest that faithful regulation and the generation of high levels of hs6871-specific mRNA depend on the presence of sequences which show homology to the 14-bp heat shock consensus element of Drosophila and, in addition, on as yet unidentified enhancer-like upstream sequences.
Keywords: heat shock, plant promoter, soybean, transgenic tobacco
Full text
PDF![1161](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2525/553914/b114cc16d831/emboj00245-0024.png)
![1162](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2525/553914/0b957e141160/emboj00245-0025.png)
![1163](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2525/553914/f711e1141e56/emboj00245-0026.png)
![1164](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2525/553914/5e29f2b5ae46/emboj00245-0027.png)
![1165](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2525/553914/46a47a1f71eb/emboj00245-0028.png)
![1166](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2525/553914/db7b0cc120e8/emboj00245-0029.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
- Bevan M. W., Mason S. E., Goelet P. Expression of tobacco mosaic virus coat protein by a cauliflower mosaic virus promoter in plants transformed by Agrobacterium. EMBO J. 1985 Aug;4(8):1921–1926. doi: 10.1002/j.1460-2075.1985.tb03871.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Corces V., Pellicer A., Axel R., Meselson M. Integration, transcription, and control of a Drosophila heat shock gene in mouse cells. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7038–7042. doi: 10.1073/pnas.78.11.7038. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Block M., Herrera-Estrella L., Van Montagu M., Schell J., Zambryski P. Expression of foreign genes in regenerated plants and in their progeny. EMBO J. 1984 Aug;3(8):1681–1689. doi: 10.1002/j.1460-2075.1984.tb02032.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Fluhr R., Kuhlemeier C., Nagy F., Chua N. H. Organ-specific and light-induced expression of plant genes. Science. 1986 May 30;232(4754):1106–1112. doi: 10.1126/science.232.4754.1106. [DOI] [PubMed] [Google Scholar]
- Gasser S. M., Laemmli U. K. Cohabitation of scaffold binding regions with upstream/enhancer elements of three developmentally regulated genes of D. melanogaster. Cell. 1986 Aug 15;46(4):521–530. doi: 10.1016/0092-8674(86)90877-9. [DOI] [PubMed] [Google Scholar]
- Herr W., Gluzman Y. Duplications of a mutated simian virus 40 enhancer restore its activity. Nature. 1985 Feb 21;313(6004):711–714. doi: 10.1038/313711a0. [DOI] [PubMed] [Google Scholar]
- Key J. L., Lin C. Y., Chen Y. M. Heat shock proteins of higher plants. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3526–3530. doi: 10.1073/pnas.78.6.3526. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Meade H. M., Long S. R., Ruvkun G. B., Brown S. E., Ausubel F. M. Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis. J Bacteriol. 1982 Jan;149(1):114–122. doi: 10.1128/jb.149.1.114-122.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagao R. T., Czarnecka E., Gurley W. B., Schöffl F., Key J. L. Genes for low-molecular-weight heat shock proteins of soybeans: sequence analysis of a multigene family. Mol Cell Biol. 1985 Dec;5(12):3417–3428. doi: 10.1128/mcb.5.12.3417. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Rochester D. E., Winer J. A., Shah D. M. The structure and expression of maize genes encoding the major heat shock protein, hsp70. EMBO J. 1986 Mar;5(3):451–458. doi: 10.1002/j.1460-2075.1986.tb04233.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schöffl F., Baumann G. Thermo-induced transcripts of a soybean heat shock gene after transfer into sunflower using a Ti plasmid vector. EMBO J. 1985 May;4(5):1119–1124. doi: 10.1002/j.1460-2075.1985.tb03748.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schöffl F., Key J. L. An analysis of mRNAs for a group of heat shock proteins of soybean using cloned cDNAs. J Mol Appl Genet. 1982;1(4):301–314. [PubMed] [Google Scholar]
- Schöffl F., Raschke E., Nagao R. T. The DNA sequence analysis of soybean heat-shock genes and identification of possible regulatory promoter elements. EMBO J. 1984 Nov;3(11):2491–2497. doi: 10.1002/j.1460-2075.1984.tb02161.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spena A., Hain R., Ziervogel U., Saedler H., Schell J. Construction of a heat-inducible gene for plants. Demonstration of heat-inducible activity of the Drosophila hsp70 promoter in plants. EMBO J. 1985 Nov;4(11):2739–2743. doi: 10.1002/j.1460-2075.1985.tb03997.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Struhl K. Naturally occurring poly(dA-dT) sequences are upstream promoter elements for constitutive transcription in yeast. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8419–8423. doi: 10.1073/pnas.82.24.8419. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Timko M. P., Kausch A. P., Castresana C., Fassler J., Herrera-Estrella L., Van den Broeck G., Van Montagu M., Schell J., Cashmore A. R. Light regulation of plant gene expression by an upstream enhancer-like element. Nature. 1985 Dec 12;318(6046):579–582. doi: 10.1038/318579a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
- Werner D., Chemla Y., Herzberg M. Isolation of poly(A)+ RNA by paper affinity chromatography. Anal Biochem. 1984 Sep;141(2):329–336. doi: 10.1016/0003-2697(84)90050-2. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]