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A tobacco protein kinase, NPK2, has a domain homologous to a domain found in activators of mitogen-activated protein kinases (MAPKKs)

Abstract

A cDNA (cNPK2) that encodes a protein of 518 amino acids was isolated from a library prepared from poly(A)+ RNAs of tobacco cells in suspension culture. The N-terminal half of the predicted NPK2 protein is similar in amino acid sequence to the catalytic domains of kinases that activate mitogen-activated protein kinases (designated here MAPKKs) from various animals and to those of yeast homologs of MAPKKs. The N-terminal domain of NPK2 was produced as a fusion protein in Escherichia coli, and the purified fusion protein was found to be capable of autophosphorylation of threonine and serine residues. These results indicate that the N-terminal domain of NPK2 has activity of a serine/threonine protein kinase. Southern blot analysis showed that genomic DNAs from various plant species, including Arabidopsis thaliana and sweet potato, hybridized strongly with cNPK2, indicating that these plants also have genes that are closely related to the gene for NPK2. The structural similarity between the catalytic domain of NPK2 and those of MAPKKs and their homologs suggests that tobacco NPK2 corresponds to MAPKKs of other organisms. Given the existence of plant homologs of an MAP kinase and tobacco NPK1, which is structurally and functionally homologous to one of the activator kinases of yeast homologs of MAPKK (MAPKKKs), it seems likely that a signal transduction pathway mediated by a protein kinase cascade that is analogous to the MAP kinase cascades proposed in yeasts and animals, is also conserved in plants.

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References

  1. Banno H, Hirano K, Nakamura T, Irie K, Nomoto S, Matsumoto K, Machida Y (1993) NPK1, a tobacco gene that encodes a protein with a domain homologous to yeast BCK1, STE11, and Byr2 protein kinases. Mol Cell Biol 13:4745–4752

  2. Banuett F (1992) Ustilago maydis, the delightful blight. Trends Genet 8:174–180

  3. Blenis J (1993) Signal transduction via the MAP kinases: Proceed at your own RSK. Proc Natl Acad Sci USA 90:5889–5892

  4. Brewster JL, de Valoir T, Dwyer ND, Winter E, Gustin MC (1993) An osmosensing signal transduction pathway in yeast. Science 259:1760–1763

  5. Brunner D, Oellers N, Szabad J, Briggs III WH, Zipursky SL, Hafen E (1994) A gain-of-function mutation in Drosophila MAP kinase activates multiple receptor tyrosine kinase signaling pathways. Cell 76:875–888

  6. Cairns BR, Ramer SW, Kornberg RD (1992) Order of action of components in the yeast pheromone response pathway revealed with a dominant allele of the STE11 kinase and the multiple pbosphorylation of the STE7 kinase. Genes Dev 6:1305–1318

  7. Chang C, Kwok SF, Bleecker AB, Meyerowitz EM (1993) Arabidopsis ethylene-response gene ETR1: Similarity of product to two-component regulators. Science 262:539–544

  8. Crews CM, Alessandrini A, Erikson RL (1992) The primary structure of MEK, a protein kinase that phosphorylates the ERK gene product. Science 258:478–480

  9. Dent P, Haser W, Haystead TAJ, Vincent LA, Roberts TM, Sturgill TW (1992) Activation of mitogen-activated protein kinase kinase by v-Raf in NIH 3T3 cells and in vitro. Science 257:1404–1407

  10. Dérijard B, Hibi M, Wu I-H, Barret T, Su B, Deng T, Karin M, Davis RJ (1994) JNK1: A protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain. Cell 76:1025–1037

  11. Duerr B, Gawienowski M, Ropp T, Jacobs T (1993) MsERKI: a mitogen-activated protein kinase from a flowering plant. Plant Cell 5:87–96

  12. Egan SE, Weinberg RA (1993) The pathway to signal achievement. Nature 365:781–783

  13. Errede B, Gartner A, Zhou Z, Nasmyth K, Ammeter G (1993) A pheromone induced kinase cascade in S. cerevisiae of the FUS3 kinase by the STE7 kinase in vitro. Nature 362:261–264

  14. Gartner A, Nasmyth K, Ammerer G (1992) Signal transduction in Saccharomyces cerevisiae requires tyrosine and threonine phosphorylation of FUS3 and KSS1. Genes Dev 6:1280–1292

  15. Gotoh Y, Matsuda S, Takenaka K, Hattori S, Iwamatsu A, Ishikawa M, Kosako H, Nishida E (1994) Characterization of recombinant Xenopus MAP kinase kinases mutated at protential phosphorylation sites. Oncogene 9:1891–1898

  16. Hanks SK, Quinn AM (1991) Protein kinase catalytic domain sequence data-base: Identification of conserved features of primary structure and classification of family members. Methods Enzymol 200:38–62

  17. Hirayama T, Oka A (1992) Novel protein kinase of Arabidopsis thaliana (APK1) that phosphorylates tyrosine, serine and threonine. Plant Mol Biol 20:653–662

  18. Howe LR, Leevers SJ, Gómez N, Nakielny S, Cohen P, Marshall CJ (1992) Activation of the MAP kinase pathway by the protein kinase raf. Cell 71:335–342

  19. Irie K, Takase MKS, Lee KS, Levin DE, Araki H, Matsumoto K, Oshima Y (1993) MKK1 and MKK2, encoding Saccharomyces cerevisiae MAP kinase-kinase homologs, function in the pathway mediated by protein kinase C. Mol Cell Biol 13:3076–3083

  20. Ito Y, Banno H, Moribe T, Hinata K, Macbida Y (1994) NPK15, a tobacco protein-serine/threonine kinase with a single hydrophobic region near the amino-terminus. Mol Gen Genet 245:1–10

  21. Jonak C, Páy A, Bögre L, Hirt H, Heberle-Bors E (1993) The plant homologue of MAP kinase is expressed in a cell cycle-dependent and organ-specific manner. Plant J 3:611–617

  22. Jonak C, Heberle-Bors E, Hirt H (1994) Map kinases: universal multi-purpose signaling tools. Plant Mol Biol 24:407–416

  23. Kato K, Matsumoto T, Koiwai A, Mizusaki S, Nishida K, Noguchi M, Tamaki E (1972) Liquid suspension culture of tobacco cells. In: Terui G (ed) Fermentation technology today. Society of Fermentation Technology Japan, Osaka, pp 689–695

  24. Kieber JJ, Rothenberg M, Roman G, Feldmann KA, Ecker JR (1993) CTR1, a negative regulator of the ethylene response pathway in Arabidopsis, encodes a member of the Raf family of protein kinases. Cell 72:427–441

  25. Kosako H, Nishida E, Gotoh Y (1993) cDNA cloning of MAP kinase kinase reveals kinase cascade pathway in yeasts to vertebrates. EMBO J 12:787–794

  26. Kyriakis JM, App H, Zhang X-f, Banerjee P, Brautigan DL, Rapp UR, Avruch J (1992) Raf-1 activates MAP kinase-kinase. Nature 358:417–421

  27. Kyriakis JM, Banerjee P, Nikolakak E, Dai T, Rubie EA, Ahmad MF, Avruch J, Woodgett JR (1994) The stress-activated protein kinase subfamily of c-Jun kinases. Nature 369:156–160

  28. Lackner MR, Kornfeld K, Miller LM, Horvitz HR, Kim SK (1994) A MAP kinase homolog, mpk-1, is involved in ras-mediated induction of vulval cell fates in Caenorhabditis elegans. Genes Dev 8:160–173

  29. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

  30. Lange-Carter CA, Pleiman CM, Gardner AM, Blumer KJ, Johnson GL (1993) A divergence in the MAP kinase regulatory network defined by MEK kinase and Raf. Science 260:315–319

  31. Lee KS, Iric K, Gotoh Y, Watanabe Y, Araki H, Nishida E, Matsumoto K, Levin DE (1993) A yeast MAP kinase homolog (MPK1) mediates signalling by protein kinase. Mol Cell Biol 13:3067–3075

  32. Maeda T, Wurgler-Murphy SM, Saito H (1994) A two-component signal transduction system that regulates the osmosensing MAP kinase cascade in yeast. Nature 369:242–245

  33. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

  34. Martin GB, Brommonschenkel SH, Chunwongse J, Frary A, Ganal MW, Spivey R, Wu T, Earle ED, Tanksley SD (1993) Map-based cloning of a protein kinase gene conferring disease resistance in tomato. Science 262:1432–1436

  35. Matsumoto S, Ito Y, Hosoi T, Takahashi Y, Machida Y (1990) Integration of Agrobacterium T-DNA into a tobacco chromosome: Possible involvement of DNA homology between T-DNA and plant DNA. Mol Gen Genet 224:309–316

  36. Mizoguchi T, Hayashida N, Yamaguchi-Shinozaki K, Kamada H, Shinozaki K (1993) ATMPKs: a gene family of plant MAP kinases in Arabidopsis thaliana. FEBS Lett 336:440–444

  37. Mizoguchi T, Gotoh Y, Nishida E, Yamaguchi-Shinozaki K, Hayashida N, Iwasaki H, Kamada H, Shinozaki K (1994) Characterization of two cDNAs that encode MAP kinases in Arabidopsis thaliana and the role of auxin in activating such kinases in plants. Plant J 5:111–122

  38. Mu J-H, Lee H-S, Kao T-h (1994) Characterization of a pollenexpressed receptor-like kinase gene of Petunia inflata and the activity of its encoded kinase. Plant Cell 6:709–721

  39. Nadin-Davis SA, Nasim A (1988) A gene which encodes a predicted protein kinase can restore some functions of the ras gene in fission yeast. EMBO J 7:985–993

  40. Neiman AM (1993) Conservation and reiteration of a kinase cascades. Trends Genet 9:390–394

  41. Onouchi H, Yokoi K, Machida C, Matsuzaki H, Oshima Y, Matsuoka K, Nakamura K, Machida Y (1991) Operation of an efficient site-specific recombination system of Zygosaccharomyces rouxii in tobacco cells. Nucleic Acids Res 19:6373–6378

  42. Ota IM, Varshavsky A (1993) A yeast protein similar to bacterial two-component regulators. Science 262:566–569

  43. Posada J, Yew N, Alm NG, Vande Woude GF, Cooper JA (1993) Mos stimulates MAP kinase in Xenopus oocytes and activates a MAP kinase kinase in vitro. Mol Cell Biol 13:2546–2553

  44. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

  45. Stafstrom JP, Altschuler M, Anderson DH (1993) Molecular cloning and expression of a MAP kinase homologue from pea. Plant Mol Biol 22:83–90

  46. Stein JC, Howlett B, Boyes DC, Nasrallah ME, Nashrallah JB (1991) Molecular cloning of a putative receptor protein kinase gene encoded at the self-incompatibility locus of Brassica oleracea. Proc Natl Acad Sci USA 88:8816–8820

  47. Stevenson BJ, Rhodes N, Errede B, Sprague Jr GF (1992) Constitutive mutants of the protein kinase STE11 activate the yeast pheromone response pathway in the absence of the G protein. Genes Dev 6:1293–1304

  48. Teague MA, Chaleff DT, Errede B (1986) Nucleotide sequence of the yeast regulatory gene STE7 predicts a protein homologous to protein kinases. Proc Natl Acad Sci USA 83:7371–7375

  49. Tsuda L, Inoue YH, Yoo M-A, Mizuno M, Hata M, Lim Y-M, Adachi-Yamada T, Ryo H, Masamune Y, Nishida Y (1993) A protein kinase similar to MAP kinase activator acts downstream of the raf kinase in Drosophila. Cell 72:407–414

  50. Walker JC, Zhang R (1990) Relationship of a putative receptor protein kinase from maize to the S-locus glycoproteins of Brassica. Nature 345:743–746

  51. Wilson C, Eller N, Gartner A, Vicente O, Harberle BE (1993) Isolation and characterization of a tobacco cDNA clone encoding a putative MAP kinase. Plant Mol Biol 23:543–551

  52. Wu Y, Han M (1994) Suppression of activated Let-60 Ras protein defines a role of Caenorhabditis elegans Sur-1 MAP kinase in vulval differentiation. Genes Dev 8:147–159

  53. Wu J, Harrison JK, Dent P, Lynch KR, Weber MJ, Sturgill TW (1993a) Identification and characterization of a new mammalian mitogen-activated protein kinase kinase, MKK2. Mol Cell Biol 13:4539–4548

  54. Wu J, Harrison JK, Vincent LA, Haystead C, Haystead TAJ, Michel H, Hunt DF, Lynch KR, Sturgill TW (1993b) Molecular structure of a protein-tyrosine/threonine kinase activating p42 mitogen-activated protein (MAP) kinase: MAP kinase kinase. Proc Natl Acad Sci USA 90:173–177

  55. Yashar BM, Kelley C, Yee K, Errede B, Zon LI (1993) Novel members of the mitogen-activated protein kinase activator family in Xenopus laevis. Mol Cell Biol 13:5738–5748

  56. Zhou Z, Gartner A, Cade R, Ammerer G, Errede B (1993) Pheromone induced signal transduction in S. cerevisiae requires the sequential function of three protein kinases. Mol Cell Biol 13:2069–2080

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Communicated by M. Sekiguchi

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Shibata, W., Banno, H., Ito, Y. et al. A tobacco protein kinase, NPK2, has a domain homologous to a domain found in activators of mitogen-activated protein kinases (MAPKKs). Molec. Gen. Genet. 246, 401–410 (1995). https://doi.org/10.1007/BF00290443

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Key words

  • Tobacco
  • Protein kinase
  • MAP kinase cascade
  • MAPKK
  • Signal transmission