Abstract
Protein kinases are important in eukaryotic signal transduction pathways. In this study we designed degenerate oligonucleotides corresponding to two conserved regions of protein kinases and using the polymerase chain reaction (PCR) have amplified a 141 bp fragment of DNA from mungbeans (Vigna radiata Rwilcz cv. Berken). Sequence analysis of the PCR products indicates that they encode several putative protein kinases with respect to their identity with other known plant protein kinases. Using one of the six fragments (CPK3-8), we isolated a 2022 bp cDNA (VrCDPK-1) from a Vigna radiata λgt11 library. VrCDPK-1 has a 96 bp 5′-untranslated region and a 465 bp 3′-untranslated region and an open reading frame of 1461 bp. VrCDPK-1 contains all of the conserved regions commonly found in calcium dependent protein kinases (CDPK). VrCDPK-1 shares 24 to 89% sequence identity with previously reported sequences for plant CDPKs at the protein level. southern analysis revealed the presence of several copies of the CDPK gene. VrCDPK-1 expression was stimulated when mungbean cuttings were treated with CaCl2, while treatment with MgCl2 had no effect. We are reporting for the first time a CDPK gene in mungbean which is inducible by mechanical strain. Cuttings treated with indole-3-acetic acid (IAA) or subjected to salt stress showed an increase in VrCDPK-1 expression. There was a dramatic stimulation in VrCDPK-1 expression 6 h after cuttings were treated with cycloheximide.
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References
Botella JR, Arteca RN: Differential expression of two calmodulin genes in response to physical and chemical stimuli. Plant Mol Biol 24: 757–766 (1994).
Botella JR, Arteca JM, Schlagnhaufer CD, Arteca RN, Phillips AT. Identification and characterization of a fulllength cDNA encoding for an auxin-induced 1-aminocy-clopropane-1-carboxylate synthase from etiolated mung bean hypocotyl segments and expression of its mRNA in response to indole-3-acetic acid. Plant Mol Biol 20: 425–436 (1992).
Botella JR, Arteca RN, Frangos JA: A mechanical strain-induced ACC synthase gene. Proc Natl Acad Sci USA 92: 1595–1598 (1995).
Braam J, Davis RW: Rain-wind-and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis. Cell 60: 357–364 (1990).
Edelman AM, Blumenthal DK, Krebs EG: Protein serine/threonine kinases. Annu Rev Biochem 56: 567–613 (1987).
Epstein E: Responses of plants to saline environments. In: Rains DW, Valentine RC, Hollaender A (eds) Genetic Engineering of Osmoregulation, pp. 7–21, Plenum Press, New York (1980).
Franco AR, Gee MA, Guilfoyle TJ: Induction and super-induction of auxin-responsive mRNAs with auxin and protein synthesis inhibitors. J Biol Chem 265: 15845–15849 (1990).
Gawienowski MC, Szymanski D, Perera IY, Zielinski: Calmodulin isoforms in Arabidopsis encoded by multiple divergent mRNAs. Plant Mol Biol 22: 215–225 (1993).
Guilfoyle TJ: Second messengers and gene expression. In: Boss WF, Morre DJ (eds) Second Messengers in Plant Growth and Development, pp. 315–326. Alan R. Liss, New York (1989).
Hanks SK, Quinn AM, Hunter T: The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science 241: 42–52 (1988).
Harper JF, Sussman MR, Schaller GE, Putman-Evans C, Charbonneau H, Harmon A: A calcium-dependent protein kinase with a regulatory domain similar to calmodulin. Science 252: 951–954 (1991).
Harper JF, Binder BM, Sussman MR: Calcium and lipid regulation of an Arabidopsis protein kinase expressed in Escherichia coli: Biochemistry 32: 3282–3290 (1993).
Hepler PK, Wayne RO: Calcium and plant development. Annu Rev Plant Physiol 36: 397–439 (1985).
Jaffe MJ, Forbes S: Thigmomorphogenesis: the effect of mechanical pertubation on plants. Plant Growth Regul 12: 313–324 (1993).
Jena PK, Reddy ASN, Poovaiah BW: Molecular cloning and sequencing of cDNA for plant calmodulin: signal-induced changes in the expression of calmodulin. Proc Natl Acad Sci USA 86: 3644–3648 (1989).
Jorgenson RA, Cuellar RE, Thompson WF: Modes and tempos in the evolution of nuclear encoded ribosomal RNA genes in legumes. Carnegie Inst Wash Yearbook 81: 98–101 (1982).
Kawasaki T, Hayashida N, Baba T, Shinozaki K, Shimada H: The gene encoding a calcium-dependent protein kinase located near the Sbe1 gene encoding a starch branching enzyme I is specifically expressed in developing rice seeds. Gene 129: 183–189 (1993).
Knight MR, Campbell AK, Smith SM, Trewavas AJ: Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors or cytoplasmatic calcium. Nature 352: 524–526 (1991).
Poovaiah BW, Reddy ASN: Calcium and signal transduction in plants. Crit Rev Plant Sci 12: 185–211 (1993).
Putman-Evans CL, Harmon AC, Cormier MJ: Purification and characterization of a novel calcium-dependent protein kinase from soybean. Biochemistry 29: 2488–2495 (1990).
Roberts DM, Harmon AC: Calcium-modulated proteins: targets of intracellular calcium signals in higher plants. Annu Rev Plant Physiol Plant Mol Biol 43: 375–414 (1990).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Sistrunk ML, Antosiewicz DM, Purugganan MM, Braam J. Arabidopsis TCH3 encodes a novel Ca2+ binding protein and shows environmentally induced and tissue specific regulation. Plant Cell 6: 1553–1565 (1994).
Suen K-L, Choi JH: Isolation and sequence of a cDNA clone for carrot calcium-dependent protein kinase: homology to calcium/calmodulin-dependent protein kinases and to calmodulin. Plant Mol Biol 17: 581–590 (1991).
Urao T, Katagiri T, Mizoguchi T, Yamaguchi-Shinozaki K, Hayashida N, Shinozaki K: Two genes that encode Ca2+-dependent protein kinases are induced by drought and high-salt stress in Arabidopsis thaliana. Mol Gen Genet 244: 331–340 (1994).
Urao T, Katagiri T, Mizoguchi T, Yamaguchi-Shinozaki K, Hayashida N, Shinozaki K: An Arabidopsis thaliana cDNA encoding Ca2+-dependent protein kinase. Plant Physiol 105: 1461–1462 (1994).
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Botella, J.R., Arteca, J.M., Somodevilla, M. et al. Calcium-dependent protein kinase gene expression in response to physical and chemical stimuli in mungbean (Vigna radiata). Plant Mol Biol 30, 1129–1137 (1996). https://doi.org/10.1007/BF00019547
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DOI: https://doi.org/10.1007/BF00019547