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Expression, tissue distribution and subcellular localization of dehydrin TAS14 in salt-stressed tomato plants

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Abstract

We previously isolated and characterized TAS14, an mRNA that is induced in tomato upon osmotic stress or abscisic acid (ABA) treatment and that shares expression and sequence characteristics with other dehydrin genes in different species. Affinity-purified antibodies against TAS14 protein were used to study the expression of TAS14 protein, both in seedlings and mature plants, its tissue distribution and its subcellular localization. TAS14 protein was not detected in 4-day-old seedlings but accumulated after ABA, NaCl or mannitol treatments. In NaCl-treated seedlings, some protein was detectable after 6 h of treatment and reached maximal levels between 24 and 48 h. Concentrations ranging from 5 to 12.5 g/l NaCl induced the protein to similar levels. In salt-stressed mature plants, TAS14 was expressed abundantly and continuously in aerial parts, but only slightly and transiently in roots. Immunocytochemical analysis of salt-treated plants showed TAS14 accumulated in adventitious root primordia and associated to the provascular and vascular tissues in stems and leaves. Immunogold electron microscopy localized TAS14 protein both in the cytosol and in the nucleus, associated to the nucleolus and euchromatin. Since TAS14 is a phosphoprotein in vivo, the classes of protein kinases potentially responsible for its in vivo phosphorylation were tested in in vitro phosphorylation assays. TAS14 protein was phosphorylated in vitro by both casein kinase II and cAMP-dependent protein kinase.

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References

  1. Alcázar A, Martín E, Lopez-Fando J, Salinas M: An improved purification procedure and properties of casein kinase II from brain. Neurochem Res 13: 829–836 (1988).

    Google Scholar 

  2. Baker J, Steel C, Dure L: Sequence and characterization of 6 Lea proteins and their genes from cotton. Plant Mol Biol 11: 277–291 (1988).

    Google Scholar 

  3. Bendayan M, Roth J, Perrelet A, Orci L: Quantitative immunocytochemical localization of pancreatic secretory proteins in subcellular compartments of the rat acinar cell. J Histochem Cytochem 28: 149–160 (1980).

    Google Scholar 

  4. Bensadoun A, Weinstein D: Assay of proteins in the presence of interfering materials. Anal Biochem 76: 241–250 (1976).

    Google Scholar 

  5. Bradford KJ, Chandler PM: Expression of ‘dehydrin-like’ proteins in embryos and seedlings of Zizania palustris and Oryza sativa during dehydration. Plant Physiol 99: 488–494 (1992).

    Google Scholar 

  6. Bray EA: Molecular responses to water deficit. Plant Physiol 103: 1035–1040 (1993).

    Google Scholar 

  7. Close TJ, Fenton RD, Moonan F: A view of plant dehydrins using antibodies specific to the carboxy terminal peptide. Plant Mol Biol 23: 279–286 (1993).

    Google Scholar 

  8. Close TJ, Kort AA, Chandler PM: A cDNA-based comparison of dehydration-induced proteins (dehydrins) in barley and corn. Plant Mol Biol 13: 95–108 (1989).

    Google Scholar 

  9. Close TJ, Lammers PJ: An osmotic stress protein of cyanobacteria is immunologically related to plant dehydrins. Plant Physiol 101: 773–779 (1993).

    Google Scholar 

  10. Dingwall C, Laskey RA: Nuclear targeting sequences — a consensus? Trends Biochem Sci 16: 478–481 (1991).

    Google Scholar 

  11. Dingwall C, Robbins J, Dilworth SM, Roberts B, Richardson WD: The nucleoplasmin nuclear location sequence is larger and more complex than that of SV40 large T-antigen. J Cell Biol 107: 841–849 (1988).

    Google Scholar 

  12. Dure LIII: The LEA proteins of higher plants. In: Verma DSP (ed) Control of Plant Gene Expression, pp. 325–335. CRC Press, Boca Raton, FL (1993).

    Google Scholar 

  13. Dure LIII, Crouch M, Harada J, Ho T-HD, Mundy J, Quatrano R, Thomas T, Sung ZR: Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol Biol 12: 475–486 (1989).

    Google Scholar 

  14. Galau GA, Close TJ: Sequences of the cotton group 2 lea/rab/dehydrin proteins encoded by lea3 cDNAs. Plant Physiol 98: 1523–1525 (1992).

    Google Scholar 

  15. Gilmour SJ, Artus NN, Thomashow MF: cDNA sequence analysis and expression of two cold-regulated genes of Arabidopsis thaliana. Plant Mol Biol 18: 13–21 (1992).

    Google Scholar 

  16. Goday A, Jensen AB, Culiáñez-Macià FA, Albà MM, Figueras M, Serratosa J, Torrent M, Pagès M: The maize abscisic acid-responsive protein Rab 17 is localized in the nucleus and cytoplasm and interacts with nuclear localization signals Plant Cell 6: 351–360 (1994).

    Google Scholar 

  17. Godoy JA, Pardo JM, Pintor-Toro JA: A tomato cDNA inducible by salt stress and abscisic acid: nucleotide sequence and expression pattern. Plant Biol 15: 695–705 (1990).

    Google Scholar 

  18. Guo W, Ward RW, Tomashow MF: Characterization of a cold-regulated wheat gene related to Arabidopsis cor47. Plant Physiol 100: 915–922 (1992).

    Google Scholar 

  19. Harlow E, Lane D: Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1988).

    Google Scholar 

  20. Hathaway GM, Traugh JA: Caseine kinase II. Meth Enzymol 99: 317–331 (1983).

    Google Scholar 

  21. Hong B, Barg R, Ho T-HD: Developmental and organ-specific expression of an ABA- and stress-induced protein in barley. Plant Mol Biol 18: 663–674 (1992).

    Google Scholar 

  22. Kalderon D, Richardson WD, Markhan AF, Smith AE: Sequence requirements for nuclear location of simian virus 40 large T antigen. Nature 311: 33–38 (1984).

    Google Scholar 

  23. Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K, Kamada H, Harada H: cDNA cloning of ECP40, an embryogenic-cell protein in carrot, and its expression during somatic and zygotic embryogenesis. Plant Mol Biol 21: 1053–1068 (1993).

    Google Scholar 

  24. Lang V, Palva ET: The expression of a rab-related gene, rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 20: 951–962 (1992).

    Google Scholar 

  25. LaRosa PC, Chen Z, Nelson DE, Singh NK, Hasegawa PM, Bressan RA. Osmotin gene expression is posttranscriptionally regulated. Plant Physiol 100: 409–415 (1992).

    Google Scholar 

  26. Mundy J, Chua NH: Abscisic acid and water-stress induce the expression of a novel rice gene. EMBO J 7: 2279–2286 (1988).

    Google Scholar 

  27. Olmsted JB: Analysis of cytoskeletal structures using blot-purified monospecific antibodies. Meth Enzymol 134: 467–472 (1986).

    Google Scholar 

  28. Pearson RB, Mitchelhill Kl, Kemp BE: Studies of protein kinase/phosphatase specificity using synthetic peptides. In: Hardie DG (ed) Protein Phosphorylation: A Practical Approach, pp. 265–291. Oxford University Press, Oxford (1993).

    Google Scholar 

  29. Perozzi G, Prakash S: RAD7 gene of Saccharomyces cerevisiae: Transcripts, nucleotide sequence analysis, and functional relationship between the RAD7 and RAD23 gene products. Mol Cell Biol 6: 1497–1507 (1986).

    Google Scholar 

  30. Piatkowski D, Schneider K, Salamini F, Bartels D: Characterization of five abscisic acid-responsive complementary DNA clones isolated from the desiccation-tolerant plant Craterostigma plantagineum and their relationship to other water-stress genes. Plant Physiol 94: 1682–1688 (1990).

    Google Scholar 

  31. Pla M, Goday A, Vilardell J, Gómez J, Pagés M: Differential regulation of ABA-induced 23–25-kDa proteins in embryo and vegetative tissues of the viviparous mutants of maize. Plant Mol Biol 13: 385–394 (1989).

    Google Scholar 

  32. Plana M, Itarte E, Goday A, Pagès M, Martinez MC: Phosphorylation of maize RAB-17 protein by casein kinase 2. J Biol Chem 266: 22510–22514 (1991).

    Google Scholar 

  33. Raikhel N: Nuclear targeting in plants. Plant Physiol 100: 1627–1632 (1992).

    Google Scholar 

  34. Rihs HP, Jans DA, Fan H, Peters R: The rate of nuclear cytoplasmic protein transport is determined by the casein kinase II site flanking the nuclear localization sequence of the SV40 T-antigen. EMBO J 10: 633–639 (1991).

    Google Scholar 

  35. Roberton M, Chandler PM: Pea dehydrins: Identification, characterisation and expression. Plant Mol Biol 19: 1031–1044 (1992).

    Google Scholar 

  36. Roberts JK, Desimone NA, Lingle WL, Dure LIII: Cellular concentrations and uniformity of cell-type accumulation of two Lea proteins in cotton embryos. Plant Cell 5: 769–780 (1993).

    Google Scholar 

  37. Roth J: Application of lectin-gold complexes for electron microscopic localization of glycoconjugates on thin sections. J Histochem Cytochem 31: 897–999 (1983).

    Google Scholar 

  38. Schmeltzer E, Krüger-Lebus S, Hahlbrock K: Temporal and spatial patterns of gene expression around sites of attempted fungal infection in parsley leaves. Plant Cell 1: 993–1001 (1989).

    Google Scholar 

  39. Schneider K, Wells B, Schmelzer E, Salamini F, Bartels D: Desiccation leads to the rapid accumulation of both cytosolic and chloroplastic proteins in the resurrection plant Craterostigma plantagineum Hothchst. Planta 189: 120–131 (1993).

    Google Scholar 

  40. Sengstag C, Hinnen A: The sequence of the Saccharomyces cerevisiae gene PHO2 codes for a regulatory protein with unusual aminoacid composition. Nucl Acids Res 15: 233–246 (1987).

    Google Scholar 

  41. Skriver K, Mundy J: Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512 (1990).

    Google Scholar 

  42. Smith DB, Johnson KS: Single-step purification of polypeptides expressed in Escherichia coli as fusion with glutathione S-transferease. Gene 67: 31–40 (1988).

    Google Scholar 

  43. Stanley KK, Luzio JP: Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human proteins. EMBO J 3: 1429–1434 (1984).

    Google Scholar 

  44. Torres-Schumann S, Godoy JA, del Pozo O, Pintor-Toro JA: Salt-induced TAS14 protein is highly phosphorylated in vivo. J Plant Physiol 139: 115–118 (1991).

    Google Scholar 

  45. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354 (1979).

    Google Scholar 

  46. Tuazon PT, Traugh JA: Casein kinase I and II- multipotential serine protein kinases: Structure, function and regulation. Adv Sec Mess Phosphoprotein Res 23: 123–164 (1991).

    Google Scholar 

  47. Vilardell J, Goday A, Freire MA, Torrent M, Martinez MC, Torne JM, Pages M: Gene sequence, developmental expression and protein phosphorylation of RAB-17 in maize. Plant Mol Biol 14: 423–432 (1990).

    Google Scholar 

  48. Wilhelmsen KC, Eggleton K, Temin HM: Nucleic acid sequences of the oncogene v-rel in reticuloendotheliosis virus strain T and its celular homolog, the proto-oncogene c-rel. J Virol 52: 172–182 (1984).

    Google Scholar 

  49. Zakut-Houri R, Bienz-Tadmor B, Givol D, Oren M: Human p53 cellular tumor antigen: cDNA sequence and expression in COS cells. EMBO J 4: 1251–1255 (1990).

    Google Scholar 

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Authors and Affiliations

  1. Instituto de Recursos Naturales y Agrobiologia, C.S.I.C., Apartado 1052, 41080, Sevilla, Spain

    José A. Godoy, Sonia Torres-Schumann, Javier Moreno & José A. Pintor-Toro

  2. Departmento de Biología Celular, Facultad de Biología, Universidad de Sevilla, 41012, Sevilla, Spain

    Rosa M. Rodrigo

Authors
  1. José A. Godoy
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  2. Rosa Lunar
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  3. Sonia Torres-Schumann
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  4. Javier Moreno
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  5. Rosa M. Rodrigo
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  6. José A. Pintor-Toro
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The first two authors contributed equally to this paper.

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Godoy, J.A., Lunar, R., Torres-Schumann, S. et al. Expression, tissue distribution and subcellular localization of dehydrin TAS14 in salt-stressed tomato plants. Plant Mol Biol 26, 1921–1934 (1994). https://doi.org/10.1007/BF00019503

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  • DOI: https://doi.org/10.1007/BF00019503

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