Abstract
Two cDNA clones, AATCDPK1 and cATCDPK2, encoding Ca2+-dependent, calmodulin-independent protein kinases (CDPK) were cloned from Arabidopsis thaliana and their nucleotide sequences were determined. Northern blot analysis indicated that the mRNAs corresponding to the ATCDPK1 and ATCDPK2 genes are rapidly induced by drought and high-salt stress but not by low-temperature stress or heat stress. Treatment of Arabidopsis plants with exogenous abscisic acid (ABA) had no effect on the induction of ATCDPK1 or ATCDPK2. These findings suggest that a change in the osmotic potential of the environment can serve as a trigger for the induction of ATCDPK1 and ATCDPK2. Putative proteins encoded by ATCDPK1 and ATCDPK2 which contain open reading frames of 1479 and 1488 bp, respectively, are designated ATCDPK1 and ATCDPK2 and show 52% identity at the amino acid sequence level. ATCDPK1 and ATCDPK2 exhibit significant similarity to a soybean CDPK (51 % and 73%, respectively). Both proteins contain a catalytic domain that is typical of serine/threonine protein kinases and a regulatory domain that is homologous to the Ca2+-binding sites of calmodulin. Genomic Southern blot analysis suggests the existence of a few additional genes that are related to ATCDPK1 and ATCDPK2 in the Arabidopsis genome. The ATCDPK2 protein expressed in Escherichia coli was found to phosphorylate casein and myelin basic protein preferentially, relative to a histone substrate, and required Ca2+ for activation.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Anderberg RJ, Walker-Simmons MK (1992) Isolation of a wheat cDNA clone for an abscisic acid-inducible transcript with homology to protein kinases. Proc Natl Acad Sci USA 89:10183–10187
Babu YS, Sack JS, Greenhough TG, Bugg CE, Means AR (1985) Three-dimensional structure of calmodulin. Nature 315:37–40
Bartels D, Schneider K, Terstappen G, Piatkowski D, Salamini F (1990) Molecular cloning of abscisic acid-modulated genes which are induced during desiccation of the resurrection plant Craterostigma plantagineum. Planta 181:27–34
Bennett MK, Kennedy MB (1987) Deduced primary structure of the β-subunit of brain type II Ca2+/calmodulin-dependent protein kinase determined by molecular cloning. Proc Natl Acad Sci USA 84:1794–1798.
Blatt MR (1990) Potassium channel currents in intact stomata] guard cells: rapid enhancement by abscisic acid. Planta 180:445–455
Braam J (1992) Regulated expression of the calmodulin-related TCH genes in cultured Arabidopsis cells: induction by calcium and heat shock. Proc Natl Acad Sci USA 89:3213–3216
Braam J, Davis RW (1990) Rain-, wind- and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis. Cell 60:357–364.
Bray EA (1988) Drought- and ABA-induced changes in polypeptide and mRNA accumulation in tomato leaves. Plant Physiol 88:1210–1214
Bray EA (1991) Regulation of gene expression by endogenous ABA during drought stress. In: Davies WJ, Jones HG (eds) Abscisic acid: physiology and biochemistry. Bios Scientific Publishers, Oxford, pp 81–98
Close TJ, Kortt AA, Chandler PM (1989) A cDNA-based comparison of dehydration-induced proteins (dehydrins) in barley and corn. Plant Mol Biol 13:95–108
Gilmour SJ, Thomashow MF (1991) Cold acclimation and coldregulated gene expression in ABA mutants of Arabidopsis thaliana. Plant Mol Biol 17:1233–1240
Gilroy S, Fricker M, Read ND, Trewavas AJ (1991) Role of calcium in signal transduction of Commelina guard cells. Plant Cell 3:333–344
Gorin MB, Yancey CB, Cline J, Revel JP, Horwitz J (1984) The major intrinsic protein (MIP) of the bovine lens fiber membrane: characterization and structure based on cDNA cloning. Cell 39:49–59
Guerrero FD, Mullet JE (1988) Reduction of turgor induces rapid changes in leaf translatable RNA. Plant Physiol 88:401–408
Guerrero FD, Jones JT, Mullet JE (1990) Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three inducible genes. Plant Mol Biol 15:11–26
Gundersen RE, Nelson DL (1987) A novel Ca2+-dependent protein kinase from Paramecium tetraurelia. J Biol Chem 262:4602–4609
Hanks SK, Quinn AM (1991) Protein kinase catalytic domain sequence database: identification of conserved features of primary structure and classification of family members. Methods Enzymol 200:38–61
Harper JF, Sussman MR, Schaller GE, Putnam-Evans C, Charbonneau H, Harmon AC (1991) A calcium-dependent protein kinase with a regulatory domain similar to calmodulin. Science 252:951–954
Harper JF, Bimder BM, Sussman MR (1993) Calcium and lipid regulation of an Arabidopsis protein kinase expressed in Escherichia coli. Biochemistry 32:3282–3290
Johnson KD, Chrispeels MJ (1992) Tonoplast-bound protein kinase phosphorylates tonoplast intrinsic protein. Plant Physiol 100:1787–1795
Johnson KD, Hofte H, Chrispeels MJ (1990) An intrinsic tonoplast protein of protein storage vacuoles in seeds is structurally related to a bacterial solute transporter (GlpF). Plant Cell 2:525–532
Kawasaki T, Hayashida N, Baba T, Shinozaki K, Shimada H (1993) A novel calcium-dependent protein kinase near the gene encoding starch branching enzyme I (sbe 1) is specifically expressed in developing rice seeds. Gene 129:183–189
Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K (1994) Cloning of cDNAs for genes that are early responsive to dehydration-stress (ERDs) in Arabidopsis thaliana L.: identification of three ERDs as lisp cognate genes. Plant Mol Biol (in press)
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lin X, Feng XH, Watson JC (1991) Differential accumulation of transcripts encoding protein kinase homologs in greening pea seedlings. Proc Natl Acad Sci USA 88:6951–6955
MacRobbie EAC (1989) Calcium influx at the plasmalemma of isolated guard cells of Commelina communis. Effect of abscisic acid. Planta 178:231–241
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Maurel C, Reizer J, Schroeder JI, Chrispeels MJ (1993) The vacuolar membrane protein γ-TIP creates water specific channels in Xenopus oocytes. EMBO J 12:2241–2247
McAinsh MR, Brownlee C, Hetherington AM (1990) Abscisic acid-induced elevation of guard cell cytosolic Ca2+ precedes stomatal closure. Nature 343:186–188
Mizoguchi T, Yamaguchi-Shinozaki K, Hayashida N, Kamada H, Shinozaki K (1993) Cloning and characterization of two cDNAs encoding casein kinase II catalytic subunits in Arabidopsis thaliana. Plant Mol Biol 21:279–289
Mundy J, Chua NH (1988) Abscisic acid and water-stress induce the expression of a novel rice gene. EMBO J 7:2270–2286
Muramatsu S, Mizuno T (1989) Nucleotide sequence of the region encompassing the glpKF operon and its upstream region containing a bent DNA sequence of Escherichia coli. Nucleic Acids Res 17:4378
Nagy F, Kay SA, Chua NH (1988) Analysis of gene expression in transgenic plants. In: Gelivin SV Schilperoort RA (eds) Plant molecular biology manual, B4. Kluwer Academic Publishers, Dordrecht, pp 1–29
Nordin K, Heino P, Palva ET (1991) Separate signal pathways regulate the expression of a low-temperature-induced gene in Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 16:1061–1071
Putnam-Evans C, Harmon AC, Cormier MJ (1990) Purification and characterization of a novel calcium-dependent protein kinase from soybean. Biochemistry 29:2488–2495
Roberts DM, Harmon AC (1992) Calcium-modulated proteins: Targets of intracellular calcium signals in higher plants. Annu Rev Plant Physiol Plant Mol Biol 43:394–414
Sandal NN, Marcker KA (1988) Soybean nodulin 26 is homologous to the major intrinsic protein of the bovine lens fiber membrane. Nucleic Acids Res 16:9347
Schaller GE, Sussman MR (1988) Phosphorylation of the plasma membrane H+-ATPase of oat roots by a calcium-stimulated protein kinase. Planta 173:509–518
Shinozaki K, Yamaguchi-Shinozaki K, Kiyosue T, Iwasaki T, Urao S (1993) Characterization of genes responsive to desiccation and their expression in Arabidopsis thaliana. In: Research in phytosynthesis, vol 4. Proceedings of the 9th International Congress on Phytosynthesis. Kluwer Academic Publishers, Dordrecht, pp 227–230
Skriver K, Mundy J (1990) Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2:503–512
Smith DB, Johnson KS (1988) Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67:31–40
Soderling TR (1990) Protein kinases. Regulation by autoinhibitory domains. J Biol Chem 265:1823–1826
Suen KL, Choi JH (1991) Isolation and sequence analysis of a cDNA clone for a carrot calcium-dependent protein kinase: homology to calcium/calmodulin-dependent protein kinases and to calmodulin. Plant Mol Biol 17:581–590
Trewavas A, Gilroy S (1991) Signal transduction in plant cells. Trends Genet 7:356–361
Tufty RM, Kretsinger RH (1975) Troponin and parvalbumin calcium binding regions predicted in myosin light chain and T4 lysozyme. Science 187:167–169
Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K (1993) An Arabidopsis myb homolog is induced by water stress and its gene product binds to the conserved MYB-recognition sequence. Plant Cell 5:1529–1539
Valvekens D, Montagu MV, Lijsebettens MV (1988). Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85:5536–5540
Van Aelst L, Hohmann S, Zimmermann FK, Jans AW, Thevelein JM (1991) A yeast homologue of the bovine lens fiber MIP gene family complements the growth defect of a Saccharomyces cerevisiae mutant on fermentable sugar but not its defect in glucose-induced RAS-mediated cAMP signalling. EMBO J 10:2095–2104
Weaver CD, Roberts DM (1992) Determination of the site of phosphorylation of nodulin 26 by the calcium-dependent protein kinase from soybean nodules. Biochemistry 31:8954–8959
Weaver CD, Crombie B, Stacey G, Roberts DM (1991) Calciumdependent phosphorylation of symbiosome membrane proteins from nitrogen-fixing soybean nodules. Plant Physiol 95:222–227
Wimmers LE, Ewing NN, Bennett AB (1992) Higher plant Ca2+-ATPase: Primary structure and regulation of mRNA abundance by salt. Proc Natl Acad Sci USA 89:9205–9209
Yamaguchi-Shinozaki K, Shinozaki K (1993) Characterization of the expression of a desiccation responsive rd29 gene of Arabidopsis thaliana and analysis of its promoter in transgenic plants. Mol Gen Genet 236:331–340
Yamaguchi-Shinozaki K, Shinozaki K (1994) A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low temperature, or high-salt stress. Plant Cell 6:251–264
Yamaguchi-Shinozaki K, Koizumi M, Urao S, Shinozaki K (1992) Molecular cloning and characterization of 9 cDNAs for genes that are responsive to desiccation in Arabidopsis thaliana: sequence analysis of one cDNA clone that encodes a putative transmembrane channel protein. Plant Cell Physiol 33:217–224
Yamamoto YT, Cheng CL, Conkling MA (1990) Root-specific genes from tobacco and Arabidopsis homologous to an evolutionarily conserved gene family of membrane channel proteins. Nucleic Acids Res 18:7449
Yoshida K, Nagano Y, Murai N, Sasaki Y (1993) Phytochromeregulated expression of the genes encoding the small GTP-binding proteins in peas. Proc Natl Acad Sci USA 90:6636–6640
Author information
Authors and Affiliations
Additional information
Communicated by M. Sekiguchi
Rights and permissions
About this article
Cite this article
Urao, T., Katagiri, T., Mizoguchi, T. et al. Two genes that encode Ca2+-dependent protein kinases are induced by drought and high-salt stresses in Arabidopsis thaliana . Molec. Gen. Genet. 244, 331–340 (1994). https://doi.org/10.1007/BF00286684
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00286684