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Characterization of a dual plant protein kinase (FsPK1) up-regulated by abscisic acid and calcium and specifically expressed in dormant seeds of Fagus sylvatica L.

Published online by Cambridge University Press:  22 February 2007

O. Lorenzo ,
C. Nicolás ,
G. Nicolás  and
D. Rodríguez
O. Lorenzo
Affiliation:
Departamento de Fisiología Vegetal, Centro Hispano-Luso de Investigaciones Agrarias, Facultad de Biología Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, E-37007, Spain
C. Nicolás
Affiliation:
Departamento de Fisiología Vegetal, Centro Hispano-Luso de Investigaciones Agrarias, Facultad de Biología Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, E-37007, Spain
G. Nicolás
Affiliation:
Departamento de Fisiología Vegetal, Centro Hispano-Luso de Investigaciones Agrarias, Facultad de Biología Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, E-37007, Spain
D. Rodríguez*
Affiliation:
Departamento de Fisiología Vegetal, Centro Hispano-Luso de Investigaciones Agrarias, Facultad de Biología Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, E-37007, Spain
*
*Correspondence Fax: +34 923 294682 E-mail: mdr@usal.es
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Abstract

An abscisic acid (ABA)-induced cDNA fragment encoding a putative protein kinase (PK) was obtained by differential screening of a cDNA library from Fagus sylvatica seeds. The full-length clone, named FsPK1, was produced by 5′ rapid amplification of cDNA ends (RACE) extension. This clone contained the 11 catalytic domains present in all protein kinases, but displayed unusual characteristics found only in a few plant PKs. FsPK1 exhibits features of both serine/threonine and tyrosine protein kinases within the catalytic domain, a putative nuclear localization signal within the regulatory domain and the consensus sequence involved in binding of 14-3-3 proteins. The catalytic domain, expressed in Escherichia coli as a fusion protein, showed Ca2+-dependent in vitro kinase activity and dual serine/threonine and tyrosine specificity. Transcription of the FsPK1 gene was reduced by seed stratification at 4°C, and clearly increased when seeds were treated with 0.1 mM ABA, correlating with the inhibition of germination. Interestingly, FsPK1 transcripts were enhanced when ABA (0.1 mM) and calcium (1 mM) were added together, while the addition of EGTA (calcium chelator) and 3,4,5,-trimethoxibenzoic acid 8-(diethylamino) octyl ester (TMB-8, a calcium antagonist) decreased its expression. Furthermore, FsPK1 transcript expression was tissue specific and accumulated only in ABA-treated seeds, but not in any ABA-treated vegetative tissues examined. These results suggest that the expression of the corresponding protein could be related to the inhibition of germination mediated by ABA in a calcium-dependent pathway.

Keywords

abscisic acidcalciumFagus sylvaticaFsPK1protein kinaseseed dormancy

Information

Type
Research Article
Information
Seed Science Research , Volume 13 , Issue 4 , December 2003 , pp. 261 - 271
Copyright
Copyright © Cambridge University Press 2003

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References

Akiyama, T., Ishida, J., Nakagawa, S., Ogawara, H., Watanabe, S., Itoh, N., Shibuya, M. and Fukami, Y. (1987) Genistein, a specific inhibitor of tyrosine-specific protein kinases. Journal of Biological Chemistry 262, 5592–5595.CrossRefGoogle ScholarPubMed
Ali, N., Halfter, U. and Chua, N.H. (1994) Cloning and biochemical characterization of a plant protein kinase that phosphorylates serine, threonine and tyrosine. Journal of Biological Chemistry 269, 31626–31629.Google Scholar
Bewley, J.D. (1997) Seed germination and dormancy. Plant Cell 9, 1055–1066.Google Scholar
Bleecker, A.B. and Schaller, G.E. (1996) The mechanism of ethylene perception. Plant Physiology 111, 653–660.Google Scholar
Botella, J.R., Arteca, J.M., Somodevilla, M. and Arteca, R.N. (1996) Calcium-dependent protein kinase gene expression in response to physical and chemical stimuli in mungbean. Plant Molecular Biology 30, 1129–1137.CrossRefGoogle ScholarPubMed
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.Google Scholar
Campalans, A., Messeguer, R., Goday, A. and Pagès, M. (1999) Plant responses to drought, from ABA signal transduction events to the action of the induced proteins. Plant Physiology and Biochemistry 37, 327–340.CrossRefGoogle Scholar
Cho, H.S., Yoon, G.M., Lee, S.S., Kim, Y.A., Hwang, I., Choi, P. and Pai, H.S. (2001) A novel dual-specificity protein kinase targeted to the chloroplast in tobacco. FEBS Letters 497, 124–130.CrossRefGoogle Scholar
Colorado, P., Nicolás, G. and Rodríguez, D. (1991) Calcium dependence of the effects of abscisic acid on RNA synthesis during germination of Cicer arietinum seeds. Physiologia Plantarum 83, 457–462.Google Scholar
Conrath, U., Silva, H. and Klessig, D.F. (1997) Protein dephosphorylation mediates salicylic acid-induced expression of PR-1 genes in tobacco. Plant Journal 11, 747–757.CrossRefGoogle Scholar
Feng, X.H., Zhao, Y., Bottino, P.J. and Kung, S.D. (1993) Cloning and characterization of a novel member of protein kinase family from soybean. Biochimica et Biophysica Acta 1172, 200–204.Google Scholar
Finkelstein, R.R., Wang, M.I., Lynch, T.J., Rao, S. and Goodman, H.M. (1998) The Arabidopsis abscisic acid response locus ABI4 encodes an APETALA2 domain protein. Plant Cell 10, 1043–1054.CrossRefGoogle ScholarPubMed
Finnie, C., Borch, J. and Collinge, D.B. (1999) 14–3–3 proteins: eukaryotic regulatory proteins with many functions. Plant Molecular Biology 40, 545–554.Google Scholar
Giraudat, J., Hauge, B.M., Valon, C., Smalle, J., Parcy, F. and Goodman, H.M. (1992) Isolation of the Arabidopsis ABI3 gene by positional cloning. Plant Cell 4, 1251–1261.Google Scholar
Gómez-Cadenas, A., Verhey, S.D., Holappa, L.D., Shen, Q., Ho, T.H.D. and Walker-Simmons, M.K. (1999) An abscisic acid induced protein kinase, PKABA1, mediates abscisic acid-suppressed gene expression in barley aleurone layers. Proceedings of the National Academy of Sciences, USA 96, 1767–1772.CrossRefGoogle ScholarPubMed
Gómez-Cadenas, A., Zentella, R., Walker-Simmons, M.K. and Ho, T.H.D. (2001) Gibberellin/abscisic acid antagonism in barley aleurone cells: site of action of the protein kinase PKABA1 in relation to gibberellin signaling molecules. Plant Cell 13, 667–679.CrossRefGoogle ScholarPubMed
Hanks, S.K., Quinn, A.M. and Hunter, T. (1988) The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science 241, 42–52.CrossRefGoogle ScholarPubMed
Hirt, H. (2000) MAP kinases in plant signal transduction. pp. 1–10in Hirt, H. (Ed.) MAP kinases in plant signal transduction. Berlin, Springer-Verlag.Google Scholar
Holappa, L.D., Walker-Simmons, M.K. (1995) The wheat abscisic acid-responsive protein kinase mRNA, PKABA1, is up-regulated by dehydration, cold temperature, and osmotic stress. Plant Physiology 108, 1203–1210.CrossRefGoogle ScholarPubMed
Koornneef, M., Reuling, G. and Karssen, C.M. (1984) The isolation and characterization of abscisic acid-insensitive mutants of Arabidopsis thaliana. Physiologia Plantarum 61, 377–383.Google Scholar
Leung, J. and Giraudat, J. (1998) Abscisic acid signal transduction. Annual Review of Plant Physiology and Plant Molecular Biology 49, 199–222.Google Scholar
Lorenzo, O., Rodríguez, D., Nicolás, G., Rodríguez, P.L. and Nicolás, C. (2001) A new protein phosphatase 2C (FsPP2C1) induced by abscisic acid is specifically expressed in dormant beechnut seeds. Plant Physiology 125, 1949–1956.Google Scholar
Lorenzo, O., Nicolás, C., Nicolás, G. and Rodríguez, D. (2002) Molecular cloning of a functional protein phosphatase 2C (FsPP2C2) with unusual features and synergistically up-regulated by ABA and calcium in dormant seeds of Fagus sylvatica. Physiologia Plantarum 114, 482–490.Google Scholar
Merlot, S., Gosti, F., Guerrier, D., Vavasseur, A. and Giraudat, J. (2001) The ABI1 and ABI2 protein phosphatases 2C act in a negative feedback regulatory loop of the abscisic acid signalling pathway. Plant Journal 25, 295–303.CrossRefGoogle Scholar
Muslin, A.J., Tanner, J.W., Allen, P.M. and Shaw, A.S. (1996) Interaction of 14–3-3 with signaling proteins is mediated by the recognition of phosphoserine. Cell 84, 889–897.Google Scholar
Nicolás, C., Nicolás, G. and Rodríguez, D. (1996) Antagonistic effects of abscisic acid and gibberellic acid on the breaking of dormancy of Fagus sylvatica seeds. Physiologia Plantarum 96, 244–250.CrossRefGoogle Scholar
Nicolás, C., Rodríguez, D., Poulsen, F., Eriksen, E.N. and Nicolás, G. (1997) The expression of an abscisic acid-responsive glycine-rich protein coincides with the level of seed dormancy in Fagus sylvatica. Plant and Cell Physiology 38, 1303–1310.Google Scholar
Nicolás, C., Nicolás, G. and Rodríguez, D. (1998) Transcripts of a gene, encoding a small GTP-binding protein from Fagus sylvatica, are induced by ABA and accumulated in the embryonic axis of dormant seeds. Plant Molecular Biology 36, 487–491.Google Scholar
Pandey, S., Tiwari, S.B., Upadhyaya, K.C. and Sopory, S.K. (2000) Calcium signaling: linking environmental signals to cellular functions. Critical Reviews in Plant Science 19, 291–318.Google Scholar
Schwartz, S.H., Léon-Kloosterziel, K.M., Koornneef, M. and Zeevart, J.A.D. (1997) Biochemical characterization of the aba2 and aba3 mutants in Arabidopsis thaliana. Plant Physiology 114, 161–166.CrossRefGoogle ScholarPubMed
Sessa, G., Raz, V., Savaldi, S. and Fluhr, R. (1996) PK12, a plant dual-specificity protein kinase of the LAMMER family, is regulated by the hormone ethylene. Plant Cell 8, 2223–2234.Google Scholar
Shen, Q., Gómez-Cadenas, A., Zhang, P., Walker-Simmons, M.K., Sheen, J. and Ho, T.H.D. (2001) Dissection of abscisic acid signal transduction pathways in barley aleurone layers. Plant Molecular Biology 47, 437–448.Google Scholar
Steinbach, H.S., Benech-Arnold, R.L., Kristof, G., Sánchez, R.A. and Marcucci-Poltri, S. (1995) Physiological basis of pre-harvest sprouting resistance in Sorghum bicolor (L.) Moench. ABA levels and sensitivity in developing embryos of sprouting resistant and susceptible varieties. Journal of Experimental Botany 46, 701–709.CrossRefGoogle Scholar
Stone, J.M. and Walker, J.C. (1995) Plant protein kinase families and signal transduction. Plant Physiology 108, 451–457.CrossRefGoogle ScholarPubMed
Thompson, J.D., Higgins, D.G. and Gibson, T.J. (1994) CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22, 4673–4680.CrossRefGoogle ScholarPubMed
Trewavas, A.J. (1988) Timing and memory processes in seed embryo dormancy. A conceptual paradigm for plant development questions. Bioessays 6, 87–92.CrossRefGoogle Scholar
Urao, T., Katagiri, T., Mizoguchi, T., Yamaguchi-Shinozaki, K., Hayashida, N. and Shinozaki, K. (1994) Two genes that encode Ca 2+ -dependent protein kinases are induced by drought and high-salt stresses in Arabidopsis thaliana. Molecular and General Genetics 244, 331–340.Google Scholar
Verhey, S.D. and Walker-Simmons, M.K. (1997) Abscisic acid mediated responses in seeds involving protein kinases and phosphatases. pp. 225–233in Ellis, R.H.;Black, M.;Murdoch, A.J.;Hong, T.D. (Eds) Basic and applied aspects of seed biology. Dordrecht, Kluwer Academic.Google Scholar
Walker-Simmons, M.K. (1987) ABA levels and sensitivity in developing wheat embryos of sprouting resistant and susceptible cultivars. Plant Physiology 84, 61–66.Google Scholar
Walker-Simmons, M.K. (1998) Protein kinases in seeds. Seed Science Research 8, 193–200.CrossRefGoogle Scholar
Wilson, C., Anglmayer, R., Vicente, O. and Heberle-Bors, E. (1995) Molecular cloning, functional expression in Escherichia coli, and characterization of multiple mitogen-activated protein kinases from tobacco. European Journal of Biochemistry 233, 249–257.Google Scholar
Wu, Y., Kuzma, J., Marechal, E., Graeff, R., Lee, H.C., Foster, R. and Chua, N.H. (1997) Abscisic acid signaling through cyclic ADP-ribose in plants. Science 278, 2126–2130.CrossRefGoogle ScholarPubMed