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Characterization of an ethylene-responsive glutathione S-transferase gene cluster in carnation

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Abstract

In this paper we present the structural analysis of two tightly linked genes from the glutathione S-transferase (GST) gene family in carnation (Dianthus caryophyllus). Southern blot analysis and restriction endonuclease mapping revealed a single cloned region of the carnation genome was highly homologous to the previously characterized ethylene-responsive GST mRNA expressed in flower petals during senescence. Nucleotide sequencing of this region revealed the presence of two tandemly arranged genes designated GST1 and GST2. Comparison of the nucleotide sequences of the cloned genomic region with the previously characterized GST cDNA clone pSR8 revealed that GST1 contains the entire transcription unit in 10 exons interrupted by 9 introns. The transcription unit of GST2 was found to be very similar to GST1 with complete conservation of intron position. In addition, the length and nucleotide sequences of the two genes' introns were highly conserved. GST2 was not completely represented by the cloned genomic region, missing the 3′ portion of the transcription unit. Primer extension analysis indicated a single transcriptional start site for transcripts which accumulate in senescing carnation petals. The 5′-flanking sequences of GST1 and GST2 were compared and regions of homology and divergence identified. These upstream sequences were compared with other plant ethylene-responsive genes and GST genes and several sequence motifs of potential importance in the regulation of GST expression were identified. A chimeric gene constructed between −1457 bp of the 5′-flanking DNA of GST1 and the coding region of β-glucuronidase was found to confer ethylene-inducible expression in flower petals following delivery of the construct into tissue by particle bombardment.

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Abbreviations

GSH:

glutathione

GST:

glutathione S-transferase

GUS:

glucuronidase

References

  1. Boller T: Ethylene in pathogenesis and disease resistance. In: Mattoo AK, Suttle JC (eds) The Plant Hormone Ethylene, pp. 293–314. CRC Press, Boca Raton, FL (1991).

    Google Scholar 

  2. Broglie KE, Biddle P, Cressman R, Broglie R: Functional analysis of DNA sequences responsible for ethylene regulation of a bean chitinase gene in transgenic tobacco. Plant Cell 1: 599–607 (1989).

    Google Scholar 

  3. Broglie R, Broglie KE: Ethylene and gene expression. In: Mattoo AK, Suttle JC (eds) The Plant Hormone Ethylene, pp. 101–113. CRC Press, Boca Raton, FL (1991).

    Google Scholar 

  4. Brown JWS: A catalogue of splice junctions and putative branch point sequences from plant introns. Nucl Acids Res 14: 9549–9559 (1986).

    Google Scholar 

  5. Cordes S, Deikman J, Margossian LJ, Fischer RL: Interaction of a developmentally regulated DNA-binding factor with sites flanking two different fruit-ripening genes from tomato. Plant Cell 1: 1025–1034 (1989).

    Google Scholar 

  6. Czarnecka E, Edelman L, Schöffl F, Key JL: Comparative analysis of physical stress responses in soybean seedlings using cloned heat shock cDNAs. Plant Mol Biol 3: 45–58 (1984).

    Google Scholar 

  7. Czarnecka E, Nagao RT, Key JE, Gurley WB: Characterization of Gmhsp 26-A, a stress gene encoding a divergent heat shock protein of soybean: heavy-metal-induced inhibition of intron processing. Mol Cell Biol 8: 1113–1122 (1988).

    Google Scholar 

  8. Deikman J, Fischer RL: Interaction of a DNA binding factor with the 5′-flanking region of an ethyleneresponsive fruit ripening gene from tomato. EMBO J 7: 3315–3320 (1988).

    Google Scholar 

  9. Dellaporta SL, Wood J, Hicks JB: A plant DNA miniprep: Version II. Plant Mol Biol Rept 1: 19–21 (1983).

    Google Scholar 

  10. Devereux P, Hacberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).

    Google Scholar 

  11. Dudler R, Hertig C, Rebmann G, Bull J, Mauch F: A pathogen-induced wheat gene encodes a protein homologous to glutathione S-transferases. Mol Plant-Microbe Interact 4: 14–18 (1991).

    Google Scholar 

  12. Friling RS, Bensimon A, Tichauer Y, Daniel V: Xenobiotic-inducible expression of murine glutathione S-transferase Ya subunit gene is controlled by an electrophile-responsive element. Proc Natl Acad Sci USA 87: 6258–6262 (1990).

    Google Scholar 

  13. Friling RS, Bergelson S, Daniel V: Two adjacent AP-1-like binding sites form the electrophile-responsive element of the murine glutathione S-transferase Ya subunit gene. Proc Natl Acad Sci USA 89: 668–672 (1992).

    Google Scholar 

  14. Grove G, Zarlengo RP, Timmerman KP, Li N-Q, Tam MF, Tu C-PD: Characterization and heterospecific expression of cDNA clones of genes in the maize GSH S-transferase multigene family. Nucl Acids Res 16: 425–438 (1988).

    Google Scholar 

  15. Hagen G, Uhrhammer N, Guilfoyle TJ: Regulation of expression of an auxin-induced soybean sequence by cadmium. J Biol Chem 263: 6442–6446 (1988).

    Google Scholar 

  16. Jefferson RA, Kavanagh TA, Bevan MW: GUS-fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).

    Google Scholar 

  17. Klein TM, Wolf ED, Wu R, Sanford JL: High-velocity microprojectiles for delivering nuclei acids into living cells. Nature 327: 70–73 (1987).

    Google Scholar 

  18. Lawton KA, Huang B, Goldsbrough PB, Woodson WR: Molecular cloning and characterization of senescence-related genes from carnation flower petals. Plant Physiol 90: 690–696 (1989).

    Google Scholar 

  19. Lawton KA, Raghothama KG, Goldsbrough PB, Woodson WR: Regulation of senescence-related gene expression in carnation flower petals by ethylene. Plant Physiol 93: 1370–1375 (1990).

    Google Scholar 

  20. Mannervik B, Danielson UH: Glutathione transferase-Structure and catalytic activity. Crit Rev Biochem Mol Biol 23: 283–337 (1988).

    Google Scholar 

  21. Meyer RC, Goldsbrough PB, Woodson WR: An ethylene-responsive flower senescence-related gene from carnation encodes a protein homologous to glutathione S-transferases. Plant Mol Biol 17: 277–281 (1991).

    Google Scholar 

  22. Moore RE, Davies MS, O'Connell KM, Harding EI, Wiegand RC, Tiemeier DC: Cloning and expression of a cDNA encoding a maize glutathione S-transferase in E. coli. Nucl Acids Res 14: 7227–7235 (1986).

    Google Scholar 

  23. Mozer TJ, Tiemeier DC, Jaworski EG: Purification and characterization of corn glutathione S-transferase. Biochemistry 22: 1068–1072 (1987).

    Google Scholar 

  24. Nguyen T, Pickett CB: Regulation of rat glutathione S-transferase Ya subunit gene expression. DNA-protein interaction at the antioxidant responsive element. J Biol Chem 267: 13535–13539 (1992).

    Google Scholar 

  25. Pickett CB, Lu AYH: Glutathione S-transferases: Gene structure, regulation, and biological function. Annu Rev Biochem 58: 743–764 (1989).

    Google Scholar 

  26. Prändl R, Kutchan TM: Nucleotide sequence of the gene for a glutathione S-transferase from cell suspensicn cultures of Silene cucubalus. Plant Physiol 99: 1729–1731 (1992).

    Google Scholar 

  27. Raghothama KG, Lawton KA, Goldsbrough PB, Woodson WR: Characterization of an ethylene-regulated flower senescence-related gene from carnation. Plant Mol Biol 17: 61–71 (1991).

    Google Scholar 

  28. Roby D, Broglie K, Gaynor J, Broglie R: Regulation of a chitinase gene promoter by ethylene and elicitors in bean protoplasts. Plant Physiol 97: 433–439 (1991).

    Google Scholar 

  29. Rushmore TH, Pickett CB: Transcriptional regulation of the rat glutathione S-transferase Ya subunit gene. Characterization of a xenobiotic-responsive element controlling inducible expression by phenolic antioxidants. J Biol Chem 265: 14648–14658 (1990).

    Google Scholar 

  30. Sanger F, Nicklen SC, Coulsen AR: DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  31. Shah DM, Hironaka CM, Wiegand RC, Harding EI, Krivi GG, Tiemeier DC: Structural analysis of a maize gene coding for glutathione S-transferase involved in herbicide detoxification. Plant Mol Biol 6: 203–211 (1986).

    Google Scholar 

  32. Takahashi Y, Yusaba M, Hiraoka Y, Nagata T: Characterization of the auxin-regulated par gene from tobacco mesophyll protoplasts. Plant J 1: 327–332 (1991).

    Google Scholar 

  33. Takahashi Y, Nagata T: ParB: An auxin-regulated gene encoding glutathione S-transferase. Proc Natl Acad Sci USA 89: 56–59 (1992).

    Google Scholar 

  34. Talalay P, DeLong MJ, Prochaska HJ: Identification of a common chemical signal regulating the induction of enzymes that protect against chemical carcinogenesis. Proc Natl Acad Sci USA 85: 8261–8265 (1988).

    Google Scholar 

  35. Telakowski-Hopkins CA, King RG, Pickett CB: Glutathione S-transferase Ya subunit gene: Identification of regulation elements required for basal level and inducible expression. Proc Natl Acad Sci USA 85: 1000–1004 (1988).

    Google Scholar 

  36. Timmerman KP: Molecular characterization of corn glutathione S-transferase isozymes involved in herbicide detoxification. Physiol Plantarum 77: 465–471 (1989).

    Google Scholar 

  37. Tsuchida S, Sato K: Glutathione transferases and cancer. Crit Rev Biochem Mol Biol 27: 337–384 (1992).

    Google Scholar 

  38. van der Zaal EJ, Droog FNJ, Boot CJM, Hensgens LAM, Hoge JHC, Schilperoort RA, Libbenga KR: Promoters of auxin-induced genes from tobacco can lead to auxin-inducible and root tip-specific expression. Plant Mol Biol 16: 983–998 (1991).

    Google Scholar 

  39. Wiegand RL, Shah DM, Mozer TJ, Harding EI, Diaz-Collier J, Saunders C, Jaworski EG, Tiemeier DC: Messenger RNA encoding a glutathione S-transferase responsible for herbicide tolerance in maize is induced in response to safener treatment. Plant Mol Biol 7: 235–243 (1986).

    Google Scholar 

  40. Yang SF, Hoffman NE: Ethylene biosynthesis and its regulation in higher plants. Annu Rev Plant Physiol 35: 155–189 (1984).

    Google Scholar 

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Authors and Affiliations

  1. Department of Horticulture, Purdue University, 47907-1165, West Lafayette, IN, USA

    Hanan Itzhaki & William R. Woodson

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  1. Hanan Itzhaki
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  2. William R. Woodson
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Itzhaki, H., Woodson, W.R. Characterization of an ethylene-responsive glutathione S-transferase gene cluster in carnation. Plant Mol Biol 22, 43–58 (1993). https://doi.org/10.1007/BF00038994

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  • DOI: https://doi.org/10.1007/BF00038994

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