Abstract
We show that a 36-base-pair-long upstream fragment from the soybean hsp17.3-B gene comprising two partly overlapping heat-shock element (HSE)-like sequences can confer heat inducibility to a reporter gene in transgenic tobacco. The heat-shock response does not display organ specificity and is not affected by light. Insertion of these HSE-like elements into the pea rbcS-3A 5' flanking fragment (position -410 to +15) either at position -410 (5' to the enhancer) or at position -49 (between the enhancer and the "TATA" box) renders the transcript level of the reporter gene light-inducible and organ-specific under heat-shock conditions. These results demonstrate the possibility of generating a unique pattern of expression (e.g., light-dependent and organ-specific heat-shock response) by artificial combination of appropriate cis-acting regulatory elements. Moreover, by using the HSE-like sequences as a weak heat-inducible enhancer in the chimeric regulatory regions we uncover the function of negative elements within the pea rbcS-3A upstream region. These negative elements are responsible for a repressed transcript level in roots as well as in dark-adapted leaves. Therefore, the upstream fragment containing two HSE-like elements can be considered a useful tool to test the function of other cis-acting elements.
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- 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]
- Amin J., Mestril R., Schiller P., Dreano M., Voellmy R. Organization of the Drosophila melanogaster hsp70 heat shock regulation unit. Mol Cell Biol. 1987 Mar;7(3):1055–1062. doi: 10.1128/mcb.7.3.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bienz M. A CCAAT box confers cell-type-specific regulation on the Xenopus hsp70 gene in oocytes. Cell. 1986 Sep 26;46(7):1037–1042. doi: 10.1016/0092-8674(86)90703-8. [DOI] [PubMed] [Google Scholar]
- Bienz M., Pelham H. R. Heat shock regulatory elements function as an inducible enhancer in the Xenopus hsp70 gene and when linked to a heterologous promoter. Cell. 1986 Jun 6;45(5):753–760. doi: 10.1016/0092-8674(86)90789-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Dente L., Cesareni G., Cortese R. pEMBL: a new family of single stranded plasmids. Nucleic Acids Res. 1983 Mar 25;11(6):1645–1655. doi: 10.1093/nar/11.6.1645. [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., Chua N. H. Developmental regulation of two genes encoding ribulose-bisphosphate carboxylase small subunit in pea and transgenic petunia plants: Phytochrome response and blue-light induction. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2358–2362. doi: 10.1073/pnas.83.8.2358. [DOI] [PMC free article] [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]
- Fluhr Robert, Moses Phyllis, Morelli Giorgio, Coruzzi Gloria, Chua Nam-Hai. Expression dynamics of the pea rbcS multigene family and organ distribution of the transcripts. EMBO J. 1986 Sep;5(9):2063–2071. doi: 10.1002/j.1460-2075.1986.tb04467.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gurley W. B., Czarnecka E., Nagao R. T., Key J. L. Upstream sequences required for efficient expression of a soybean heat shock gene. Mol Cell Biol. 1986 Feb;6(2):559–565. doi: 10.1128/mcb.6.2.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirel B., Bouet C., King B., Layzell D., Jacobs F., Verma D. P. Glutamine synthetase genes are regulated by ammonia provided externally or by symbiotic nitrogen fixation. EMBO J. 1987 May;6(5):1167–1171. doi: 10.1002/j.1460-2075.1987.tb02350.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karin M., Haslinger A., Holtgreve H., Richards R. I., Krauter P., Westphal H. M., Beato M. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature. 1984 Apr 5;308(5959):513–519. doi: 10.1038/308513a0. [DOI] [PubMed] [Google Scholar]
- Kirk M. M., Kirk D. L. Translational regulation of protein synthesis, in response to light, at a critical stage of Volvox development. Cell. 1985 Jun;41(2):419–428. doi: 10.1016/s0092-8674(85)80015-5. [DOI] [PubMed] [Google Scholar]
- Klemenz R., Gehring W. J. Sequence requirement for expression of the Drosophila melanogaster heat shock protein hsp22 gene during heat shock and normal development. Mol Cell Biol. 1986 Jun;6(6):2011–2019. doi: 10.1128/mcb.6.6.2011. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuhlemeier C., Fluhr R., Green P. J., Chua N. H. Sequences in the pea rbcS-3A gene have homology to constitutive mammalian enhancers but function as negative regulatory elements. Genes Dev. 1987 May;1(3):247–255. doi: 10.1101/gad.1.3.247. [DOI] [PubMed] [Google Scholar]
- 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]
- McKnight S. L., Kingsbury R. C., Spence A., Smith M. The distal transcription signals of the herpesvirus tk gene share a common hexanucleotide control sequence. Cell. 1984 May;37(1):253–262. doi: 10.1016/0092-8674(84)90321-0. [DOI] [PubMed] [Google Scholar]
- Mestril R., Rungger D., Schiller P., Voellmy R. Identification of a sequence element in the promoter of the Drosophila melanogaster hsp23 gene that is required for its heat activation. EMBO J. 1985 Nov;4(11):2971–2976. doi: 10.1002/j.1460-2075.1985.tb04031.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mestril R., Schiller P., Amin J., Klapper H., Ananthan J., Voellmy R. Heat shock and ecdysterone activation of the Drosophila melanogaster hsp23 gene; a sequence element implied in developmental regulation. EMBO J. 1986 Jul;5(7):1667–1673. doi: 10.1002/j.1460-2075.1986.tb04410.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagy F., Kay S. A., Boutry M., Hsu M. Y., Chua N. H. Phytochrome-controlled expression of a wheat Cab gene in transgenic tobacco seedlings. EMBO J. 1986 Jun;5(6):1119–1124. doi: 10.1002/j.1460-2075.1986.tb04335.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Odell J. T., Nagy F., Chua N. H. Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. 1985 Feb 28-Mar 6Nature. 313(6005):810–812. doi: 10.1038/313810a0. [DOI] [PubMed] [Google Scholar]
- Parker C. S., Topol J. A Drosophila RNA polymerase II transcription factor contains a promoter-region-specific DNA-binding activity. Cell. 1984 Feb;36(2):357–369. doi: 10.1016/0092-8674(84)90229-0. [DOI] [PubMed] [Google Scholar]
- Pauli D., Spierer A., Tissières A. Several hundred base pairs upstream of Drosophila hsp23 and 26 genes are required for their heat induction in transformed flies. EMBO J. 1986 Apr;5(4):755–761. doi: 10.1002/j.1460-2075.1986.tb04278.x. [DOI] [PMC free article] [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]
- 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]
- Riddihough G., Pelham H. R. Activation of the Drosophila hsp27 promoter by heat shock and by ecdysone involves independent and remote regulatory sequences. EMBO J. 1986 Jul;5(7):1653–1658. doi: 10.1002/j.1460-2075.1986.tb04408.x. [DOI] [PMC free article] [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]
- Shermoen A. W., Jongens J., Barnett S. W., Flynn K., Beckendorf S. K. Developmental regulation by an enhancer from the Sgs-4 gene of Drosophila. EMBO J. 1987 Jan;6(1):207–214. doi: 10.1002/j.1460-2075.1987.tb04740.x. [DOI] [PMC free article] [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. The 5' ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature. 1980 Aug 28;286(5776):854–860. doi: 10.1038/286854a0. [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]