Abstract
In higher plants, as-1-like cis elements mediate auxin- and salicylic acid-inducible transcription. Originally found in viral and T-DNA promoters, they are also functional elements of plant promoters activated during the defence response against pathogens. Tobacco bZIP transcription factor TGA1a was the first recombinant protein shown to bind to as-1. cDNAs for two novel tobacco as-1-binding bZIP proteins (TGA2.1 and TGA2.2) were isolated. Revealing a high degree of amino acid identity in the bZIP domain (89%) and the C-terminus (79%), the two TGA2 factors differ remarkably with respect to the length of the N-terminus (170 amino acids in TGA2.1 versus 43 amino acids in TGA2.2). TGA2.1 and TGA2.2, but not TGA1a, interacted with ankyrin repeat protein NPR1, a central activator of the plant defence response. In contrast, TGA2.1 and TGA1a, but not TGA2.2, functioned as transcriptional activators in yeast. Apart from conferring transcriptional activation, the N-terminal domain of TGA2.1 led to reduced in vitro as-1-binding activity and almost completely abolished binding to one half site of this bifunctional element. When being part of a heterodimer with TGA2.2, TGA2.1 was efficiently recruited to a single half site, though double occupancy of the element was still preferred. In contrast, TGA1a preferred to bind to only one palindrome, a feature that was also maintained in heterodimers between TGA1a and TGA2.1 or TGA2.2.
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Alexander, D., Goodman, R.M., Gut-Rella, M., Glascock, C., Weymann, K., Friedrich, L., Maddox, D., Ahl-Goy, P., Luntz, T., Ward, E. et al. 1993. Increased tolerance to two oomycete pathogens in transgenic tobacco expressing pathogenesis-related protein 1a. Proc. Natl. Acad. Sci. USA 90: 7327–7331.
Benfey, P.N., Ren, L. and Chua, N.-H. 1989. The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J. 8: 2195–2202.
Braun, H.P., Emmermann, M., Mentzel, H. and Schmitz, U.K. 1994. Primary structure and expression of a genes encoding the cytosolic ribosomal protein S4 from potato. Biochim. Biophys. Acta 1218: 435–438.
Cao, H., Glazebrook, J., Clarke, J.D., Volko, S. and Dong, S. 1997. The Arabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats. Cell 88: 57–63.
Chuang, C.F., Running, M.P., Williams, R.W. and Meyerowitz, E.M. 1999. The PERIANTHIA gene encodes a bZIP protein involved in the determination of floral organ number in Arabidopsis thaliana. Genes Dev. 13: 334–344.
Corpet F. 1988. Multiple sequence alignment with hierarchical clustering. Nucl. Acids Res. 16: 10881–10890.
Dong, X. 1998. SA, JA, ethylene, and disease resistance in plants. Curr. Opin. Plant Biol. 1: 316–323.
Droog, F.N.J., Spek, A., van der Kooy, A., der Ruyter, A., Hoge, H., Libbenga, K., Hooykaas, P. and van der Zaal, B. 1995. Promoter analysis of the auxin-regulated tobacco glutathione S-transferase genes Nt103-1 and Nt103-35. Plant Mol. Biol. 29: 413–429.
Feinberg, A.P. and Vogelstein, B. 1983. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132: 6–13.
Fourney, R., Miyakoshi, J., Day III, R.S. and Paterson, M.C. 1988. Northern blotting: efficient RNA staining and transfer. Focus 10: 5–7.
Fromm, H., Katagiri, F. and Chua, N.H. 1989. An octopine synthase enhancer element directs tissue-specific expression and binds ASF-1, a factor from tobacco nuclear extracts. Plant Cell 1: 977–984.
Fromm, H., Katagiri, F. and Chua, N.H. 1991. The tobacco transcription activator TGA1a binds to a sequence in the 50 upstream region of a gene encoding a TGA1a-related protein. Mol. Gen. Genet. 229: 181–188.
Gaffney, J., Friedrich, L., Vernoolj, B., Negrotto, D., Nye, G., Uknes, S., Ward, E., Kessmann, H. and Ryals, J. 1993. Requirement of salicylic acid for the induction of systemic acquired resistance. Science 261: 754–756.
James, P., Halladay, J. and Craig, E.A 1996. Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144: 1425–1436.
Katagiri, F., Lam, E. and Chua, N.-H. 1989. Two tobacco DNAbinding proteins with homology to the nuclear factor CREB. Nature 340: 727–730.
Kawata, T., Imada, T., Shiraishi, H., Okada, K., Shimura, Y. and Ivabuchi, M. 1992. A DNA clone encoding an HBP-1b homologue in Arabidopsis thaliana. Nucl. Acids Res. 20: 1141–1147.
Kozak, M. 1981. Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes. Nucl. Acids Res. 24: 5233–5262.
Kusaba, M., Takahashi, Y. and Nagata, T. 1996. A multiplestimuli-responsive as-1-related element of parA gene confers responsiveness to cadmium but not to copper. Plant Physiol. 111: 1161–1167.
Laemmli U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Lam, E. and Lam, Y. K.-P. 1995. Binding site requirements and differential representation of TGA factors in nuclear ASF-1 activity. Nucl. Acids Res. 23: 3778–3785.
Liu, X.J. and Lam, E. 1994. Two binding sites for the plant transcription factor ASF-1 can respond to auxin treatments in transgenic tobacco. J. Biol. Chem. 269: 668–675.
Malamy, J., Carr, J.P., Klessig, D.F. and Raskin, I. 1990. Salicylic acid: a likely endogenous signal in the resistance response of tobacco to tobacco mosaic virus. Science 250: 1002–1004.
Menkens, A.E., Schindler, U. and Cashmore, A.R. 1995. The Gbox: a ubiquitous regulatory DNA element in plants bound by the GBF family of bZIP proteins. Trends Biochem. Sci. 20: 506–510.
Miao, Z.-H., Liu, X. and Lam, E. 1994. TGA3 is a distinct member of the TGA family of bZIP transcription factors in Arabidopsis thaliana. Plant Mol. Biol. 25: 1–11.
Neuhaus, G., Neuhaus-Url, G., Katagiri, F., Seipel, K. and Chua, N.-H. 1994. Tissue-specific expression of as-1 in transgenic tobacco. Plant Cell. 6: 827–834.
Niggeweg, R. and Gatz, C. 1997. Isolation of TGA2.1 (accession No. U90214), a Member of a new subclass of the TGA family of bZIP transcription factors in Nicotiana tabacum. Plant Physiol. 113: 1464.
Pascuzzi, P., Hamilton, D., Bodily, K. and Arias, J. 1998. Auxin-induced stress potetiates trans-activation by a conserved basic/leucine-zipper factor. J. Biol. Chem. 273: 26631–26637.
Prat, S., Willmitzer, L. and Sanchez-Serrano, J.J. 1989. Nuclear proteins binding to a cauliflower mosaic virus 35S truncated promoter. Mol. Gen. Genet. 17: 209–214.
Qin, X.-F., Holuigue, L. Horvath, D.M. and Chua, N.-H. 1994. Immediate early transcription activation by salicylic acid via the cauliflower mosaic virus as-1 element. Plant Cell 6: 863–874.
Rieping, M., Fritz, M., Prat, S. and Gatz, C. 1994. A dominant negative mutant of PG13 suppresses transcription from a cauliflower mosaic virus 35S truncated promoter in transgenic tobacco plants. Plant Cell 6: 1087–1098.
Ryals, J., Weymann, K., Lawton, K., Friedrich, L., Ellis, D., Steiner, H.Y., Johnson, J., Delaney, T.P., Jesse, T., Vos, P. and Uknes, S. 1997. The Arabidopsis NIM1 protein shows homology to the mammalian transcription factor inhibitor I kappa B. Plant Cell 9: 425–439.
Schindler, U., Beckmann, H. and Cashmore, A.R. 1992. TGA1 and G-box binding factors: two distinct classes of Arabidopsis leucine zipper proteins compete for the G-box-like element TGACGTGG. Plant Cell 4: 1309–1319.
Strompen, G., Gruner, R. and Pfitzner, U.M. 1998. An as-1-like motif controls the level of expression of the gene for the pathogenesis-related protein 1a from tobacco. Plant Mol. Biol. 37: 871–883.
Takahashi, Y., Kuroda, H., Tanaka, T., Machida, Y., Takebe, I. and Nagata T. 1989. Isolation of an auxin-regulated gene cDNA expressed during the transition from G0 to S phase in tobacco mesophyll protoplasts. Proc. Natl. Acad. Sci. USA 86: 9279–9283.
Tokuhisa, J.B., Singh, K., Dennis, E.S. and Peacock, W.J. 1990. A DNA binding protein factor recognizes two binding domains within the octopine synthase enhancer element. Plant Cell 2: 215–224.
Ulmasov, T., Hagen, G. and Guilfoyle, T. 1994. The ocs element in the soybean GH2/4 promoter is activated by both active and inactive auxin and salicylic acid analogues. Plant Mol. Biol. 26: 1055–1064.
Ward, E.R., Uknes, S.J., Williams, S.C., Dincher, S.S., Wiederhold, D.L., Alexander, D.C., Ahl-Goy, P., Métraux, J.-P. and Ryals, J.A. 1991. Coordinate gene activity in response to agents that induce systemic acquired resistance. Plant Cell 3: 1085–1094.
Xiang, C., Miao, Z. and Lam, E. 1995. Isolation of TGA6, a new member of the TGA family of bZIP transcription factors in Arabidopsis thaliana. Plant Physiol. 109, 721.
Xiang, C., Miao, Z. and Lam, E. 1996. Coordinate activation of as-1-type elements and a tobacco glutathione S-transferase gene by auxins, salicylic acid, methyl-jasmonate and hydrogen peroxide. Plant Mol. Biol. 32: 415–426.
Xiang, C., Miao, Z. and Lam, E. 1997. DNA-binding properties, genomic organization and expression pattern of TGA6, a new member of the TGA family of bZIP transcription factors in Arabidopsis thaliana. Plant Mol. Biol. 34: 403–415.
Zhang, B., Foley, R.C. and Singh, K.B. 1993. Isolation and characterization of two related Arabidopsis ocs-element bZIP binding proteins. Plant J. 4: 711–716.
Zhang, B. and Singh, K.B. 1994. ocs element promoter sequences are activated by auxin and salicylic acid in Arabidopsis. Proc. Natl. Acad. Sci. USA 91: 2507–2511.
Zhang, S. and Klessig, D.F. 1997. Salicylic acid activates a 48-kD MAP kinase in tobacco. Plant Cell 9: 809–824.
Zhang, Y., Fan, W., Kinkema, M., Li, X. and Dong, X. 1999. Interaction of NPR1 with basic leucine zipper protein transcription factors that bind sequences required for salicylic acid induction of the PR-1 gene. Proc. Natl. Acad. Sci. USA 96: 6523–6528.
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Niggeweg, R., Thurow, C., Weigel, R. et al. Tobacco TGA factors differ with respect to interaction with NPR1, activation potential and DNA-binding properties. Plant Mol Biol 42, 775–788 (2000). https://doi.org/10.1023/A:1006319113205
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DOI: https://doi.org/10.1023/A:1006319113205