Abstract
Recently, a novel type of calcium-dependent protein kinase (CDPK) that requires neither calmodulin nor phospholipids for activation, has been described in plants. We have isolated a cDNA clone for carrot CDPK by probing a library of somatic embryo cDNAs with oligonucleotides corresponding to highly conserved regions of protein kinases. The product of this gene overexpressed in Escherichia coli reacted strongly with monoclonal antibodies to soybean CDPK. The deduced amino acid sequence of carrot CDPK reveals two major functional domains. An N-terminal catalytic domain with greatest homology to calcium/calmodulin-dependent protein kinase type II from rat brain is coupled to a C-terminal calcium-binding domain resembling calmodulin. These features of the primary sequence explain how CDPK binds calcium and suggest a model for CDPK regulation based on similarities to animal calcium/calmodulin-dependent protein kinases.
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Battey, NH, Venis, MA: Calcium-dependent protein kinase from apple fruit membranes is calmodulin-independent but has calmodulin-like properties. Planta 176: 91–97 (1988).
Biermann, B, Johnson, EM, Feldman, LJ: Characterization and distribution of a maize cDNA encoding a peptide similar to the catalytic region of second messenger dependent protein kinases. Plant Physiol 94: 1609–1615 (1990).
Blowers, DP, Boss, WF, Trewavas, AJ: Rapid changes in plasma membrane protein phosphorylation during initiation of cell wall digestion. Plant Physiol 86: 505–509 (1988).
Bogre, L, Olah, Z, Dudits, D: Ca2+-dependent protein kinase from alfalfa (Medicago varia): partial purification and autophosphorylation. Plant Sci 85: 135–144 (1988).
Borkird, C, Choi, JH, Jin, Z-H, Franz, G, Hatzopoulos, P, Chorneau, R, Bonas, U, Pelegri, F, Sung, ZR: Developmental regulation of embryonic genes in plants. Proc Natl Acad Sci USA 85: 6399–6403 (1988).
Budde, RJA, Randall, DD: Light as a signal influencing the phosphorylation status of plant proteins. Plant Physiol 94: 1501–1504 (1990).
Cawley, KC, Ramachandran, C, Gorin, FA, Walsh, DA: Nucleotide sequence of cDNA encoding the catalytic subunit of phosphorylase kinase from rat soleus muscle. Nucl Acids Res 16: 2355–2356 (1988).
Chiatante, D, Levi, M, Sgorbati, S, Sparvoli, E: Variations of phosphorylation of protein membrane components in pea root meristems during germination. J Plant Growth Regul 8: 107–119 (1989).
Choi, JH, Liu, L-S, Borkird, C, Sung, ZR: Cloning of genes developmentally regulated during plant embryogenesis. Proc Natl Acad Sci USA 84: 1906–1910 (1987).
Colbran, RJ, Smith, KM, Schworer, CM, Fong, Y-L, Soderling, TR: Regulatory domain of calcium/calmodulin-dependent protein kinase II: mechanism of inhibition and regulation by phosphorylation. J Biol Chem 264: 4800–4804 (1989).
Crum, CJ, Hu, J, Hiddinga, HJ, Roth, DA: Tobacco mosaic virus infection stimulates the phosphorylation of a plant protein associated with double-stranded RNA-dependent protein kinase activity. J Biol Chem 263: 13440–13443 (1988).
Davis, TN, Urdea, MS, Masiarz, FR, Thorner, J: Isolation of the yeast calmodulin gene: calmodulin is an essential protein. Cell 47: 423–431 (1986).
Grab, D, Feger, M, Ebel, J: An endogenous factor from soybean (Glycine max L.) cell cultures activates phosphorylation of a protein which is dephosphorylated in vivo in elicitor-challenged cells. Planta 179: 340–348 (1989).
Guerriero, V, Jr, Russo, MA, Olson, NJ, Putkey, JA, Means, AR: Domain organization of chicken gizzard myosin light chain kinase deduced from a cloned cDNA. Biochemistry 25: 8372–8381 (1986).
Hanks, SK: Homology probing: identification of cDNA clones encoding members of the protein-serine kinase family. Proc Natl Acad Sci USA 84: 388–392 (1987).
Hanks, SK, Quinn, AM, Hunter, T: The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science 241: 42–52 (1988).
Hanley, RM, Means, AR, Ono, T, Kemp, BE, Burgin, KE, Waxham, N, Kelly, PT: Functional analysis of a complementary DNA for the 50-kilodalton subunit of calmodulin kinase. Science 237: 293–297 (1987).
Harmon, AC, Putnam-Evans, C, Cormier, MJ: A calcium-dependent but calmodulin-independent protein kinase from soybean. Plant Physiol 83: 830–837 (1987).
Hiddinga, HJ, Crum, CJ, Hu, J, Roth, DA: Viroid-induced phosphorylation of a host protein related to a dsRNA-dependent protein kinase. Science 241: 451–453 (1988).
Hunter, T: A thousand and one protein kinases. Cell 50: 823–829 (1987).
Huynh, TV, Young, RA, Davis, RW: Constructing and screening cDNA libraries in lambda gt10 and lambda gt11. In: Glover, DM (ed) DNA Cloning: A Practical Guide, pp. 49–78, IRL Press, Oxford, England (1985).
Kelly, PT, Weinberger, RP, Waxham, MN: Active sitedirected inhibition of Ca/calmodulin-dependent protein kinase type II by a bifunctional calmodulin-binding peptide. Proc Natl Acad Sci USA 85: 4991–4995 (1988).
Klucis, E, Polya, GM: Localization, solubilization and characterization of plant membrane-associated calcium-dependent protein kinases. Plant Physiol 88: 164–171 (1988).
Koritsas, VM: Effect of ethylene and ethylene precursors on protein phosphorylation and xylogenesis in tuber explants of Helianthus tuberosus L. J Exp Bot 39: 375–386 (1988).
Krishnan, HB, Pueppke, SG: Heat shock triggers rapid protein phosphorylation in soybean seedlings. Biochem Biophys Res Commun 148: 762–767 (1987).
Kyo, M, Harada, H: Specific phosphoproteins in the initial period of tobacco pollen embryogenesis. Planta 182: 58–63 (1990).
Lawton, MA, Yamamoto, RT, Hanks, SK, Lamb, CJ: Molecular cloning of plant transcripts encoding protein kinase homologs. Proc Natl Acad Sci USA 86: 3140–3144 (1989).
Lin, CR, Kapiloff, MS, Durgerian, S, Tatemoto, K, Russo, AF, Hanson, P, Schulman, H, Rosenfeld, MG: Molecular cloning of a brain-specific calcium/calmodulin-dependent protein kinase. Proc Natl Acad Sci USA 84: 5962–5966 (1987).
Ling, V, Zielinski, RE: Cloning of cDNA sequences encoding the calcium-binding protein, calmodulin, from barley (Hordeum vulgare L.). Plant Physiol 90: 714–719 (1989).
Lukas, TJ, Wiggins, ME, Watterson, DM: Amino acid sequence of a novel calmodulin from the unicellular alga Chlamydomonas. Plant Physiol 78: 477–483 (1985).
Martiny-Baron, G, Scherer, GFE: Phospholipid-stimulated protein kinase in plants. J Biol Chem 264: 18052–18059 (1989).
Ohtsuka, E, Matsuki, S, Ikehara, M, Takahashi, Y, Matsubara, K: An alternative approach to deoxyoligonucleotides as hybridization probes by an insertion of deoxyinosine at ambiguous codon positions. J Biol Chem 260: 2605–2608 (1985).
Pearson, RB, Wettenhall, REH, Means, AR, Hartshorne, DJ, Kemp, BE: Autoregulation of enzymes by pseudosubstrate prototypes: myosin light chain kinase. Science 241: 970–973 (1988).
Pearson, RB, Woodgett, JR, Cohen, P, Kemp, BE: Substrate specificity of a multifunctional calmodulin-dependent protein kinase. J Biol Chem 260: 14471–14476 (1985).
Poovaiah, BW, Reddy, ASN: Calcium messenger system in plants. CRC Crit Rev Plant Sci 6: 47–103 (1987).
Putnam-Evans, CL, Harmon, AC, Cormier, MJ: Purification and characterization of a novel calcium-dependent protein kinase from soybean. Biochemistry 29: 2488–2495 (1990).
Putnam-Evans, C, Harmon, AC, Palevitz, BA, Fecheimer, M, Cormier, MJ: Calcium-dependent protein kinase is localized with F-actin in plant cells. Cell Motil Cytoskel 12: 12–22 (1989).
Ranjeva, R, Boudet, AM: Phosphorylation of proteins in plants: regulatory effects and potential involvement in stimulus/response coupling. Ann Rev Plant Physiol 38: 73–93 (1987).
Rogers, SO, Bendich, AJ: Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol Biol 5: 69–76 (1985).
Sambrook, J, Fritsch, EF, Maniatis, T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor NY (1989).
Sasagawa, T, Ericsson, LH, Walsh, KA, Schreiber, WE, Fischer, EH, Titani, K: Complete amino acid sequence of human brain calmodulin. Biochemistry 21: 2565–2569 (1982).
Short, TW, Briggs, WR: Characterization of a rapid, blue light-mediated change in detectable phosphorylation of a plasma membrane protein from etiolated pea (Pisum sativum L.) seedlings. Plant Physiol 92: 179–185 (1990).
Takahashi, Y, Kato, K, Hayashizaki, Y, Wakabayashi, T, Ohtsuka, E, Matsuki, S, Ikehara, M, Matsubara, K: Molecular cloning of the human cholecystokinin gene by use of a synthetic probe containing deoxyinosine. Proc Natl Acad Sci USA 82: 1931–1935 (1985).
Walker, JC, Zhang, R: Relationship of a putative receptor-protein kinase from maize to the S-locus glycoproteins of Brassica. Nature 345: 743–746 (1990).
Wood, WJ, Gitschier, J, Lasky, LA, Lawn, RM: Base composition-independent hybridization in tetramethylammonium chloride: a method for oligonucleotide screening of highly complex gene libraries. Proc Natl Acad Sci USA 82: 1585–1588 (1985).
Wong, Y-S, McMichael, RW, Jr, Lagarias, JC: Properties of a polycation-stimulated protein kinase associated with purified Avena phytochrome. Plant Physiol 91: 709–718 (1989).
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Suen, KL., Choi, J.H. 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 (1991). https://doi.org/10.1007/BF00037045
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DOI: https://doi.org/10.1007/BF00037045