Plant Molecular Biology

, Volume 59, Issue 5, pp 713–722 | Cite as

ZmMPK6, a Novel Maize MAP Kinase that Interacts with 14-3-3 Proteins

  • Marco Lalle
  • Sabina Visconti
  • Mauro Marra
  • Lorenzo Camoni
  • Riccardo Velasco
  • Patrizia Aducci


Although an increasing body of evidence indicates that plant MAP kinases are involved in a number of cellular processes, such as cell cycle regulation and cellular response to abiotic stresses, hormones and pathogen attack, very little is known about their biochemical properties and regulation mechanism. In this paper we report on the identification and characterization of a novel member of the MAP kinase family from maize, ZmMPK6. The amino acid sequence reveals a high degree of identity with group D plant MAP kinases. Recombinant ZmMPK6, expressed in Escherichia coli, is an active enzyme able to autophosphorylate. Remarkably, ZmMPK6 interacts in vitro with GF14-6, a maize 14-3-3 protein and the interaction is dependent on autophosphorylation. The interacting domain of ZmMPK6 is on the C-terminus and is comprised between amino acid 337 and amino acid 467. Our results represent the first evidence of an interaction between a plant MAP kinase and a 14-3-3 protein. Possible functional roles of this association in vivo are discussed.


14-3-3 protein MAP kinase protein-protein interaction Zea mays 



Extracellular-signal Regulated Kinase


glutathione S-transferase


mitogen-activated protein kinase


mitogen-activated protein kinase kinase kinase


Mitogen-activated protein kinase/Extracellular signal-regulated kinase Kinase)


Maize H+-ATPase isoform 2


polyacrylamide gel electrophoresis


polyvinylidene fluoride


reverse transcriptase polimerase chain reaction


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  1. Aducci, P., Camoni, L., Marra, M., Visconti, S. 2002From cytosol to organelles: 14-3-3 proteins as multifunctional regulators of plant cellIUBMB Life534955PubMedGoogle Scholar
  2. Berberich, T., Sano, H., Kusano, T. 1999Involvement of a MAP kinase, ZmMPK5, in senescence and recovery from␣low-temperature stress in maizeMol. Gen. Genet.262534542CrossRefPubMedGoogle Scholar
  3. Bhat, RA, Riehl, M., Santandrea, G., Velasco, R., Slocombe, S., Donn, G., Steinbiss, H.H., Thompson, R.D., Becker, H.A. 2003Alteration of GCN5 levels in maize reveals dynamic responses to manipulating histone acetylationPlant J.33455469CrossRefPubMedGoogle Scholar
  4. Bradford, M.M. 1976A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein-dye bindingAnal. Biochem.72248254PubMedGoogle Scholar
  5. Calderini, O., Bogre, L., Vicente, O., Binarova, P., Heberle-Bors, E., Wilson, C. 1998A novel tobacco mitogen-activated protein (MAP) kinase kinase, NtMEK1, activates the cell cycle-regulated p43Ntf6 MAP kinaseJ. Cell Sci.11130913100PubMedGoogle Scholar
  6. Cheong, Y.H., Moon, B.C., Kim, J.K., Kim, C.Y., Kim, I.H., Park, C.Y., Kim, J.C., Park, B.O., Koo, S.C., Yoon, H.W., Chung, W.S., Lim, C.O., Lee, S.Y., Cho, M.J. 2003Plant Physiol.13219611972CrossRefPubMedGoogle Scholar
  7. Clark-Lewis, I., Sanghera, J.S., Pelech, S.L. 1991Definition of a consensus sequence for peptide substrate recognition by p44mpk, the meiosis-activated myelin basic protein kinaseJ. Biol. Chem.2661518015184PubMedGoogle Scholar
  8. Cobb, M.H., Goldsmith, E.J. 1995How MAP kinases are regulatedJ. Biol. Chem.2701484314846CrossRefPubMedGoogle Scholar
  9. Davis, R.J. 2000Signal transduction by the JNK group of MAP kinasesCell103239252CrossRefPubMedGoogle Scholar
  10. Fanger, G.R., Widmann, C., Porter, A.C., Sather, S., Johnson, G.L., Vaillancourt, R.R 199814-3-3 proteins interact with specific MEK kinasesJ. Biol. Chem.27334763483CrossRefPubMedGoogle Scholar
  11. Finnie, C., Borch, J., Collinge, D.B. 199914-3-3 proteins: eukaryotic regulatory proteins with many functionsPlant Mol. Biol.40545554CrossRefPubMedGoogle Scholar
  12. Fu, H., Subramanian, R.R., Masters, S.C. 200014-3-3 proteins: structure, function and regulationAnnu. Rev. Pharmacol. Toxicol.40617647CrossRefPubMedGoogle Scholar
  13. Fuglsang, A.T., Visconti, S., Drumm, K., Jahn, T., Stensballe, A., Mattei, B., Jensen, O.N., Aducci, P., Palmgren, M.G. 1999Binding of 14-3-3 protein to the plasma membrane H+-ATPase AHA2 involves the three Cterminal residues Tyr(946)-Thr-Val and requires phosphorylation of Thr(947)J. Biol. Chem.2743677436780CrossRefPubMedGoogle Scholar
  14. Fullone, M.R., Visconti, S., Marra, M., Fogliano, V., Aducci, P. 1998Fusicoccin effect on the in vitro interaction between plant 14-3-3 proteins and plasma membrane H+-ATPase J. Biol. Chem.27376987702CrossRefPubMedGoogle Scholar
  15. Gupta, R., Huang, Y., Kieber, J., Luan, S. 1998Identification of a dual specificity protein phosphatase that inactivates a MAP kinase from ArabidopsisPlant J.16581589CrossRefPubMedGoogle Scholar
  16. Huang, Y., Li, H., Gupta, R., Morris, P.C., Luan, S., Kieber, J.J. 2000ATMPK4, an Arabidopsis homolog of mitogen-activated protein kinase, is activated in vitro by AtMEK1 through threonine phosphorylationPlant Physiol.12213011310CrossRefPubMedGoogle Scholar
  17. Ichimura, K., Mizoguchi, T., Yoshida, R., Yuasa, T., Shinozaki, K. 2000Various abiotic stresses rapidly activate Arabidopsis MAP kinases ATMPK4 and ATMPK6Plant J.24655665CrossRefPubMedGoogle Scholar
  18. Ichimura, K., Shinozaki, K., Tena, G., Sheen, J., Henry, H., Champion, A., Kreis, M., Zhang, S., Hirt, H., Wilson, C., Heberle-Bors, E., Ellis, B.E., Morris, P.C., Innes, R.W., Ecker, J.R., Scheel, D., Klessig, D.F., Machida, Y., Mundi, J., Ohashi, Y., Walker, J.C. 2002Mitogen-activated protein kinase cascades in plants: a new nomenclatureTrends Plant Sci.7301308CrossRefPubMedGoogle Scholar
  19. Igarashi, D., Ishida, S., Fukazawa, J., Takashi, Y. 200114-3-3 proteins regulate intracellular localization of the bZIP␣transcriptional activator RSGPlant Cell1324832497CrossRefPubMedGoogle Scholar
  20. Jonak, C., Kiegerl, S., Ligterink, W., Barker, P.J., Huskinsson, N.S., Hirt, H. 1996Stress signaling in plants: a mitogen-activated protein kinase pathway is activated by cold and droughtProc. Natl. Acad. Sci. USA931127411279CrossRefPubMedGoogle Scholar
  21. Jonak, C., Okresz, L., Bogre, L., Hirt, H. 2002Complexity, cross talk and integration of plant MAP kinase signallingCurr. Opin. Plant Biol.5415424CrossRefPubMedGoogle Scholar
  22. Kieber, J.J., Rothenberg, M., Roman, G., Feldmann, K.A., Ecker, J.R. 1993CTR1, a negative regulator of the ethylene response pathway in Arabidopsis, encodes a member of the raf family of protein kinasesCell72427441CrossRefPubMedGoogle Scholar
  23. Ku, N.O., Liao, J., Omary, M.B. 1998Phosphorylation of human keratin 18 serine 33 regulates binding to 14-3-3 proteinsEMBO J.1718921906CrossRefPubMedGoogle Scholar
  24. Kultz, D. 1998Phylogenetic and functional classification of mitogen and stress-activated protein kinasesJ. Mol. Evol.46571588PubMedGoogle Scholar
  25. Laemmli, U.K. 1970Cleavage of the structural proteins during the assembly of the head of the bacteriophage T4Nature277680685CrossRefGoogle Scholar
  26. Liu, Y.C., Liu, Y., Elli, C., Yoshida, H., Lipkowitz, S., Altman, A. 1997Serine phosphorylation of Cbl induced by phorbol ester enhances its association with 14-3-3 proteins in T cells via a novel serine-rich 14-3-3-binding motifJ. Biol. Chem.27299799985CrossRefPubMedGoogle Scholar
  27. Mockaitis, K., Howell, S.H. 2000Auxin induces mitogenic activated protein kinase (MAPK) activation in roots of Arabidopsis seedlingsPlant J.24785796CrossRefPubMedGoogle Scholar
  28. Muslin, A.J., Tanner, J.W., Allen, P.M., Shaw, A.S. 1996Interaction of 14-3-3 with signaling proteins is mediated by the recognition of phosphoserineCell84889897CrossRefPubMedGoogle Scholar
  29. Ouaked, F., Rozhon, W., Lecourieux, D., Hirt, H. 2003A MAPK pathway mediates ethylene signaling in plantsEMBO J.2212821288CrossRefPubMedGoogle Scholar
  30. Pearson, W.R. 1990Rapid and sensitive sequence comparison␣with FASTP and FASTAMethod. Enzymol.1836398Google Scholar
  31. Roberts, R.L., Mosch, H.U., Fink, G.R. 199714-3-3 proteins are essential for RAS/MAPK cascade signaling during pseudohyphal development in S. cerevisiaeCell8910551065CrossRefPubMedGoogle Scholar
  32. Robinson, M.J., Cobb, M.H. 1997Mitogen-activated protein kinase pathwaysCurr. Opin. Cell Biol.9180186CrossRefPubMedGoogle Scholar
  33. Song, F., Goodman, R.M. 2002OsBIMK1, a rice MAP kinase gene involved in disease resistance responsesPlanta2159971005CrossRefPubMedGoogle Scholar
  34. Tena, G., Asai, T., Chiu, W.L., Sheen, J. 2001Plant mitogen-activated protein kinase signaling cascadesCurr. Opin. Plant Biol.4392400CrossRefPubMedGoogle Scholar
  35. Thompson, J.D., Higgins, D.G., Gibson, T.J. 1994CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choiceNucl. Acids Res.2246734680PubMedGoogle Scholar
  36. Tzivion, G., Avruch, J. 200214-3-3 proteins: active cofactors in cellular regulation by serine/threonine phosphorylationJ. Biol. Chem.27730613064CrossRefPubMedGoogle Scholar
  37. Hemert, M.J., Steensma, H.Y., Heusden, G.P.H. 200114-3-3 proteins: key regulator of cell division, signalling and apoptosisBioEssays23936946CrossRefPubMedGoogle Scholar
  38. Visconti, S., Camoni, L., Fullone Lalle, M.R. M., Marra, M., Aducci, P. 2003Mutational analysis of the interaction between 14-3-3 proteins and plant plasma membrane H+-ATPaseJ. Biol. Chem.27881728178CrossRefPubMedGoogle Scholar
  39. Weston, C.R., Davis, R.J. 2002The JNK signal transduction pathwayCurr. Opin. Genet. Dev.121421CrossRefPubMedGoogle Scholar
  40. Xing, H., Zhang, S., Weinheimer, C., Kovacs, A., Muslin, A.J. 200014-3-3 proteins block apoptosis and differentially regulate MAPK cascadesEMBO J.19349358CrossRefPubMedGoogle Scholar
  41. Yaffe, M.B., Rittinger, K., Volinia, S., Caron, P.R., Aitken, A., Leffers, H., Gamblin, S.J., Smerdon, S.J., Cantley, L.C. 1997The structural basis for 14-3-3: phosphopeptide binding specificityCell91961971CrossRefPubMedGoogle Scholar
  42. Zhang, S., Klessig, D.F. 2001MAPK cascades in plant defense signalingTrends Plant Sci.6520527CrossRefPubMedGoogle Scholar
  43. Zheng, Q., Yin, G., Yan, C., Cavet, M., Berk, B.C. 200414-3-3beta binds to big mitogen-activated protein kinase 1 (BMK1/ERK5) and regulates BMK1 functionJ. Biol. Chem.27987878791CrossRefPubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Marco Lalle
    • 1
    • 2
  • Sabina Visconti
    • 1
  • Mauro Marra
    • 3
  • Lorenzo Camoni
    • 1
  • Riccardo Velasco
    • 4
    • 5
  • Patrizia Aducci
    • 1
  1. 1.Department of BiologyUniversity of Rome “Tor Vergata”RomeItaly
  2. 2.Department of Infectious, Parasitic and Immuno-mediated DiseasesIstituto Superiore di Sanita’RomeItaly
  3. 3.Department of Biological and Environmental StudiesUniversity of SannioBeneventoItaly
  4. 4.Max Planck Institut fuer ZuechtungsforschungKoelnGermany
  5. 5.Istituto Agrario San Michele all’AdigeSan Michele a/AdigeItaly

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