Artículo

Estamos trabajando para incorporar este artículo al repositorio
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

We isolated a full-length cDNA clone (StCDPK1) encoding a calcium-dependent protein kinase (CDPK) by screening a stolon tip cDNA library from potato plants (Solarium tuberosum L.). The predicted amino acid sequence of the cDNA reveals a high degree of similarity with other members of the CDPK family except in the N-terminal region. As described for other CDPKs, StCDPK1 has a putative N-terminal myristoylation sequence. A coupled transcription/translation system was used to demonstrate that this post-translational modification occurs in vitro. The behaviour of the myristoylated form of StCDPK1 during its purification on a phenyl-Sepharose column mimics that of the endogenous potato enzyme suggesting that this modification occurs in vivo. In addition, a possible palmitoylation site is present in StCDPK1. Southern blot analysis suggests that more than one CDPK isoform is present in potato plants. Northern blot analysis of steady-state mRNA levels for StCDPK1 in different tissues of potato plants shows that the transcript is differentially expressed in tuberizing stolons. The transcript appears in the early steps of tuber formation before the induction of other genes, such as Pin2 and patatin. This result parallels previous data on CDPK activity in potato plants which was highest at the beginning of tuberization. Our results suggest that StCDPK1 is developmentally regulated. The early and transient expression of this CDPK isoform in the tuberization process suggests that this kinase could trigger a cascade of phosphorylation events involved in tuber induction.

Registro:

Documento: Artículo
Título:Molecular characterization of StCDPK1, a calcium-dependent protein kinase from Solanum tuberosum that is induced at the onset of tuber development
Autor:Raiíces, M.; Chico, J.M.; Téllez-Iñón, M.T.; Ulloa, R.M.
Filiación:Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, CONICET, Acultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina
Palabras clave:Calcium-dependent protein kinases; Differential expression; N-terminal myristoylation; Solanum tuberosum; StCDPK1; Tuberization; amino terminal sequence; cDNA clone; chemical modification; DNA library; enzyme activity; gene induction; genetic transcription; growth regulation; homology; northern blotting; plant development; potato tuber; protein kinase C; Amino acids; DNA; Enzymes; Phosphorylation; Plant cell culture; Amino Acid Sequence; Blotting, Northern; Blotting, Southern; Calcium-Binding Proteins; Cloning, Molecular; DNA, Complementary; DNA, Plant; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Plant; Molecular Sequence Data; Myristic Acid; Plant Proteins; Protein Kinases; RNA, Messenger; RNA, Plant; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Solanum tuberosum; Eukaryota; Solanum tuberosum; Solarium; Tuberosum
Año:2001
Volumen:46
Número:5
Página de inicio:591
Página de fin:601
DOI: http://dx.doi.org/10.1023/A:1010661304980
Título revista:Plant Molecular Biology
Título revista abreviado:Plant. Mol. Biol.
ISSN:01674412
CAS:Calcium-Binding Proteins; DNA, Complementary; DNA, Plant; Myristic Acid, 544-63-8; Plant Proteins; Protein Kinases, EC 2.7.1.37; RNA, Messenger; RNA, Plant; StCDPK1 protein, Solanum tuberosum, EC 2.7.1.-
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674412_v46_n5_p591_Raiices

Referencias:

  • Alwine, J.C., Kemp, D.J., Stark, G.R., Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes (1977) Proc. Natl. Acad. Sci. USA, 74, pp. 5350-5354
  • Balamani, V., Veluthambi, K., Poovaiah, B.W., Effect of calcium on tuberization in potato (1986) Plant Physiol., 80, pp. 856-858
  • Bonner, W.M., Laskey, R.A., A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels (1974) Eur. J. Biochem., 46, pp. 83-88
  • Breviario, D., Morello, L., Giani, S., Molecular cloning of two novel rice cDNA sequences encoding putative calcium-dependent protein kinases (1995) Plant Mol. Biol., 27, pp. 953-967
  • Bush, D.S., Calcium regulation in plant cells and its role in signaling (1995) Annu. Rev. Plant Physiol. Plant Mol. Biol., 46, pp. 95-122
  • Church, G.M., Gilbert, W., Genomic sequencing (1984) Proc. Natl. Acad. Sci. USA, 81, pp. 1991-1995
  • Dilworth, D.D., McCarrey, J.R., Single-step elimination of contaminating DNA prior to reverse transcriptase PCR (1992) PCR Meth. Appl., 1, pp. 279-282
  • Ellard-Ivey, M., Hopkins, R.B., White, T.J., Lomax, T.L., Cloning, expression and N-terminal myristoylation of CpCPK1, a calcium-dependent protein kinase from zucchini (Cucurbita pepo L.) (1999) Plant Mol. Biol., 39, pp. 199-208
  • Ewing, E.E., The role of hormones in potato (Solanum tuberosum L.) tuberization (1987) Plant Hormones and their Role in Plant Growth and Development, pp. 515-538. , P.J. Davies (Ed.), Kluwer Academic Publishers, Dordrecht, Netherlands
  • Harmon, A.C., Gribskov, M., Harper, J.F., CDPKs: A kinase for every Ca2+ signal? (2000) Trends Plant Sci., 5, pp. 154-159
  • Harper, J.F., Sussman, M.R., Schaller, G.E., Putnam-Evans, C., Charbonneau, H., Harmon, A.C., A calcium-dependent protein kinase with a regulatory domain similar to calmodulin (1991) Science, 252, pp. 951-954
  • Hrabak, E.M., Dickmann, L.J., Satterlee, J.S., Sussman, M.R., Characterisation of eight new members of the calmodulin-like domain protein kinase gene family from Arabidopsis thaliana (1996) Plant Mol. Biol., 31, pp. 405-412
  • Jackson, S.D., Multiple signaling pathways control tuber induction in potato (1999) Plant Physiol., 119, pp. 1-8
  • Jackson, S.D., Sonnewald, U., Willmitzer, L., Characterisation of a gene that is expressed in leaves at higher levels upon tuberization in potato and upon flowering in tobacco (1993) Planta, 189, pp. 593-596
  • Jackson, S., Gascón, J., Carrera, E., Monte, E., Prat, S., Cloning and expression analysis of a gene that shows developmental regulation upon tuberization in potato (1997) Plant Mol. Biol., 33, pp. 169-174
  • Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685
  • Lakatos, L., Hutvagner, G., Banfalvi, Z., Potato protein kinase StCPK1: A putative evolutionary link between CDPKs and CRKs (1998) Biochim. Biophys. Acta, 1442, pp. 101-108
  • MacIntosh, G.C., Ulloa, R.M., Raíces, M., Téllez-Iñón, M.T., Changes in calcium-dependent protein kinase activity during in vitro tuberization in potato (1996) Plant Physiol., 112, pp. 1541-1550
  • Marks, F., (1996) Protein Phosphorylation, , VCH, Weinheim/New York
  • Martin, M.L., Busconi, L., Membrane localization of a rice calcium-dependent protein kinase (CDPK) is mediated by myristoylation and palmitoylation (2000) Plant J., 24, pp. 429-435
  • McCabe, J.B., Berthiaume, L.G., Functional roles for fatty acylated amino-terminal domains in subcellular localization (1999) Mol. Biol. Cell, 10, pp. 3771-3786
  • Mochly-Rosen, D., Localization of protein kinases by anchoring proteins: A theme in signal transduction (1995) Science, 268, pp. 247-251
  • Murashige, T., Skoog, F., A revised medium for rapid growth and bio assays with tobacco tissue cultures (1962) Physiol. Plant., 15, pp. 473-497
  • Murray, M.G., Thompson, W.F., Rapid isolation of high molecular weight plant DNA (1980) Nucl. Acids Res., 8, pp. 4321-4325
  • Poovaiah, B.W., Reddy, A.S.N., Calcium and signal transduction in plants (1993) Crit. Rev. Plant Sci., 12, pp. 185-211
  • Prat, S., Frommer, W.B., Hofgen, R., Keil, M., Kossmann, J., Koster-Topfer, M., Liu, X.J., Willmitzer, L., Gene expression during tuber development in potato plants (1990) FEBS Lett., 268, pp. 334-338
  • Roberts, D.M., Harmon, A.C., Calcium-modulated proteins: Targets of intracellular calcium signals in higher plants (1992) Annu. Rev. Plant Physiol. Plant Mol. Biol., 43, pp. 375-414
  • Short, J.M., Fernandez, J.M., Sorge, J.A., Huse, W.D., Lambda ZAP: A bacteriophage lambda expression vector with in vivo excision properties (1988) Nucl. Acids Res., 16, pp. 7583-7600
  • Sopory, S.K., Munshi, M., Protein kinases and phosphatases and their role in cellular signaling in plants (1998) Crit. Rev. Plant Sci., 17, pp. 245-318
  • Taylor, M.A., Kumar, A., George, L.A., Davies, H.V., Isolation and molecular characterisation of a tuberization-related cDNA clone from potato (Solanum tuberosum L.) (1992) Plant Cell Rep., 11, pp. 623-626
  • Taylor, M.A., Mad Arif, S.A., Pearce, S.R., Davies, H.V., Kumar, A., George, L.A., Differential expression and sequence analysis of ribosomal protein genes induced in stolon tips of potato (Solanum tuberosum L.) during the early stages of tuberization (1992) Plant Physiol., 100, pp. 1171-1176
  • Taylor, M.A., Burch, L.R., Davies, H.V., Changes in polyamine biosynthesis during the initial stages of tuberization in potato (Solanum tuberosum L.) (1993) Plant Physiol., 141, pp. 370-372
  • Taylor, M.A., George, L.A., Ross, H.A., Davies, H.V., cDNA cloning and characterisation of an α-glucosidase gene from potato (Solanum tuberosum L.) (1998) Plant J., 13, pp. 419-425
  • Thompson, G.A., Okuyama, H., Lipid-linked proteins of plants (2000) Prog. Lipid Res., 39, pp. 19-39
  • Ulloa, R.M., Torres, H.N., Ochatt, C.M., Téllez-Iñón, M.T., Ca2+ calmodulin-dependent protein kinase activity in the ascomycetes Neurospora crassa (1991) Mol. Cell. Biochem., 102, pp. 155-163
  • Ulloa, R.M., MacIntosh, G.C., Melchiorre, M., Mentaberry, A.N., Dallari, P., Moriconi, D.N., Téllez-Iñón, M.T., Protein kinase activity in different stages of potato (Solanum tuberosum L.) microtuberization (1997) Plant Cell Rep., 16, pp. 426-429
  • Vosberg, H.-P., The polymerase chain reaction: An improved method for the analysis of nucleic acids (1989) Humangenetik, 83, pp. 1-15

Citas:

---------- APA ----------
Raiíces, M., Chico, J.M., Téllez-Iñón, M.T. & Ulloa, R.M. (2001) . Molecular characterization of StCDPK1, a calcium-dependent protein kinase from Solanum tuberosum that is induced at the onset of tuber development. Plant Molecular Biology, 46(5), 591-601.
http://dx.doi.org/10.1023/A:1010661304980
---------- CHICAGO ----------
Raiíces, M., Chico, J.M., Téllez-Iñón, M.T., Ulloa, R.M. "Molecular characterization of StCDPK1, a calcium-dependent protein kinase from Solanum tuberosum that is induced at the onset of tuber development" . Plant Molecular Biology 46, no. 5 (2001) : 591-601.
http://dx.doi.org/10.1023/A:1010661304980
---------- MLA ----------
Raiíces, M., Chico, J.M., Téllez-Iñón, M.T., Ulloa, R.M. "Molecular characterization of StCDPK1, a calcium-dependent protein kinase from Solanum tuberosum that is induced at the onset of tuber development" . Plant Molecular Biology, vol. 46, no. 5, 2001, pp. 591-601.
http://dx.doi.org/10.1023/A:1010661304980
---------- VANCOUVER ----------
Raiíces, M., Chico, J.M., Téllez-Iñón, M.T., Ulloa, R.M. Molecular characterization of StCDPK1, a calcium-dependent protein kinase from Solanum tuberosum that is induced at the onset of tuber development. Plant. Mol. Biol. 2001;46(5):591-601.
http://dx.doi.org/10.1023/A:1010661304980