Skip to main content
Log in

Do 14-3-3 proteins and plasma membrane H+-ATPases interact in the barley epidermis in response to the barley powdery mildew fungus?

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

14-3-3 proteins form a family of highly conserved proteins with central roles in many eukaryotic signalling networks. In plants, they bind to and activate the plasma membrane H+-ATPase, creating a binding site for the phytotoxin fusicoccin. Barley 14-3-3 transcripts accumulate in the epidermis upon inoculation with the powdery mildew fungus. We have isolated a cDNA encoding a plasma membrane H+-ATPase (HvHA1), which is also induced by powdery mildew attack. The C-terminal domain of this H+-ATPase interacts with 14-3-3 proteins in the yeast two-hybrid system. Inoculation with the powdery mildew fungus, or treatment with fusicoccin, results in an increase in fusicoccin binding ability of barley leaf membranes. Overlay assays show a fungus-induced increase in binding of digoxygenin-labelled 14-3-3 protein to several proteins including a 100 kDa membrane protein, probably the plasma membrane H+-ATPase. These effects are seen specifically in the inoculated epidermis and not in the whole leaf. We propose that 14-3-3 proteins are involved in an epidermis-specific response to the powdery mildew fungus, possibly via an activation of the plasma membrane H+-ATPase.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Baunsgaard, L., Fuglsang, A.T., Jahn, T., Korthout, H.A.A.J., de Boer, A.H. and Palmgren, M.G. 1998. The 14–3–3 protein associates with the plasma membrane H+-ATPase to generate a fusicoccin binding complex and a fusicoccin responsive system. Plant J. 13: 661–671.

    Google Scholar 

  • Booij, P.P., Roberts, M.R., Vogelzang, S.A., Kraayenhof, R. and de Boer, A.H. 1999. 14–3–3 proteins double the number of outwardrectifying K+ channels available for activation in tomato cells. Plant J. 20: 673–683.

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal. Biochem. 72: 248–254.

    Google Scholar 

  • Brandt, J., Thordal-Christensen, H., Vad, K., Gregersen, P.L. and Collinge, D.B. 1992. A pathogen induced gene of barley encodes a protein showing high similarity to a protein kinase regulator. Plant J. 2: 815–820.

    Google Scholar 

  • Durfee, T., Becherer, K., Chen, P.L., Yeh, S.H., Yang, Y.Z., Kilburn, A.E., Lee, W.H. and Elledge, S.J. 1993. The retinoblastoma protein associates with the protein phosphatase type-1 catalytic subunit. Genes Dev. 7: 555–569.

    Google Scholar 

  • Felix, G. and Boller, T. 1995. Systemin induces rapid ion fluxes and ethylene biosynthesis in Lycopersicon peruvianum cells. Plant J. 7: 381–389

    Google Scholar 

  • Finnie, C., Borch, J. and Collinge, D.B. 1999. 14–3–3 proteins: eukaryotic regulatory proteins with many functions. Plant Mol. Biol. 40: 545–554.

    Google Scholar 

  • Fuglsang, A. T., Visconti, S., Drumm, K., Jahn, T., Stensballe, A., Mattei, B., Jensen, O.N., Aducci, P. and Palmgren, M. G. 1999. Binding of 14–3–3 protein to the plasma membrane H+-ATPase AHA2 involves the three C-terminal residues Tyr946–Thr-Val and requires phosphorylation of Thr947. J. Biol. Chem. 274: 36774–36780.

    Google Scholar 

  • Gianinazzi-Pearson, V., Arnould, C., Oufattole, M., Arango, M. and Gianinazzi, S. 2000. Differential activation of H+-ATPase genes by an arbuscular mycorrhizal fungus in root cells of transgenic tobacco. Planta 211: 609–613.

    Google Scholar 

  • Gregersen, P. L., Thordal-Christensen, H., Förster, H. and Collinge, D.B. 1997. Differential gene transcript accumulation in barley leaf epidermis and mesophyll in response to attack by Blumeria graminis f.sp. hordei. Physiol. Mol. Plant Path. 51: 85–97.

    Google Scholar 

  • Hess, W.R., Golz, R. and Börner, T. 1998. Analysis of randomly selected cDNAs reveals the expression of stress-and defencerelated genes in the barley mutant albostrians. Plant Sci. 133: 191–201.

    Google Scholar 

  • Jahn, T., Fuglsang, A.T., Olsson, A., Brüntrup, I.M,. Collinge, D.B., Volkmann, D., Sommarin, M., Palmgren, M.G. and Larsson, C. 1997. The 14–3–3 protein interacts directly with the C-terminal region of the plant plasma membrane H+-ATPase. Plant Cell 9: 1805–1814.

    Google Scholar 

  • Kippert, F. 1995. A rapid permeabilization procedure for accurate quantitative determination of β-galactosidase activity in yeast cells. FEMS Microbiol. Lett. 128: 201–206.

    Google Scholar 

  • Kølster, P., Munk, L., Stølen, O. and Løhde, J. 1986. Near isogenic barley lines with genes for resistance to powdery mildew. Crop Sci. 26: 903–907.

    Google Scholar 

  • Larsson, C., Sommarin, M. and Widell, S. 1994. Isolation of highly purified plant plasma membranes and separation of inside-out and right-side-out vesicles. Meth. Enzymol. 228: 451–469.

    Google Scholar 

  • Marra, M., Olivari, C., Visconti, S., Albumi, C., Aducci, P. and De Michelis, M.I. 2000. A phosphopeptide corresponding to the cytosolic stretch connecting transmembrane segments 8 and 9 of the plasma membrane H+-ATPase binds 14–3–3 proteins and inhibits fusicoccin-induced activation of the H+-ATPase. Plant Biol. 2: 11–16.

    Google Scholar 

  • Moorhead, G., Douglas, P., Cotelle, V., Harthill, J., Morrice, N., Meek, S., Deiting, U., Stitt, M., Scarabel, M., Aitken, A. and Mackintosh, C. 1999. Phosphorylation-dependent interactions between enzymes of plant metabolism and 14–3–3 proteins. Plant J. 18: 1–12.

    Google Scholar 

  • Murphy, P.J., Langridge, P. and Smith, S.E. 1997. Cloning plant genes differentially expressed during colonization of roots of Hordeum vulgare by the vesicular-arbuscular mycorrhizal fungus Glomus intraradices. New Phytol. 135: 291–301.

    Google Scholar 

  • Oecking, C. and Hagemann, K. 1999. Association of 14–3–3 proteins with the C-terminal autoinhibitory domain of the plant plasmamembrane H+-ATPase generates a fusicoccin-binding complex. Planta 207: 480–482.

    Google Scholar 

  • Oecking, C., Piotrowski, M., Hagemeier, J. and Hagemann, K. 1997. Topology and target interaction of the fusicoccin-binding 14–3–3 homologs of Commelina communis. Plant J. 12: 441–453.

    Google Scholar 

  • Olsson, A., Svennelid, F., Ek, B., Sommarin, M. and Larsson, C. 1998. A phosphothreonine residue at the C-terminal end of the plasma membrane H+-ATPase is protected by fusicoccininduced 14–3–3 binding. Plant Physiol. 118: 551–555.

    Google Scholar 

  • Piotrowski, M., Morsomme, P., Boutry, M. and Oecking, C. 1998. Complementation of the Saccharomyces cerevisiae plasma membrane H+-ATPase by a plant H+-ATPase generates a highly abundant fusicoccin binding site. J. Biol. Chem. 273: 30018–30023.

    Google Scholar 

  • Roberts, M.R. and Bowles, D.J. 1999. Fusicoccin, 14–3–3 proteins, and defence responses in tomato plants. Plant Physiol. 119: 1243–1250.

    Google Scholar 

  • Schaller, A. and Oecking, C. 1999. Modulation of plasma membrane H+-ATPase activity differentially activates wound and pathogen defense responses in tomato plants. Plant Cell 11: 263–272.

    Google Scholar 

  • Seehaus, K. and Tenhaken, R. 1998. Cloning of genes by mRNA differential display induced during the hypersensitive reaction of soybean after inoculation with Pseudomonas syringae pv. glycinea. Plant Mol. Biol. 38: 1225–1234.

    Google Scholar 

  • Sutton, P.N., Henry, M.J. and Hall, J.L. 1999. Glucose, and not sucrose, is transported from wheat to wheat powdery mildew. Planta 208: 426–430.

    Google Scholar 

  • Thordal-Christensen, H. and Smedegaard-Petersen, V. 1988. Comparison of resistance-inducing abilities of virulent and avirulent races of Erysiphe graminis f.sp. hordei and a race of Erysiphe graminis f.sp. tritici. Plant Path. 37: 20–27.

    Google Scholar 

  • Thordal-Christensen, H., Brandt, J., Cho, B.H., Gregersen, P.L., Rasmussen, S.K., Smedegaard-Petersen, V. and Collinge, D.B. 1992. cDNA cloning and characterization of two barley peroxidase transcripts induced differentially by the powdery mildew fungus. Physiol. Mol. Plant Path. 40: 395–409.

    Google Scholar 

  • Thordal-Christensen H., Gregersen, P.L. and Collinge, D.B. 2000. The barley/Blumeria (syn. Erysiphe) graminis interaction: a case study. In: A.J. Slusarenko, R.S.S. Fraser and L.C. van Loon (Eds.) Mechanisms of Resistance to Plant Diseases, Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 77–100.

    Google Scholar 

  • Thorpe, G.G. and Kricka, L.J. 1986. Enhanced chemiluminescent reactions catalyzed by horseradish peroxidase. Meth. Enzymol. 133: 331–353.

    Google Scholar 

  • Toroser, D., Athwal, G.S. and Huber S.C. 1998. Site-specific regulatory interaction between spinach leaf sucrose-phosphate synthase and 14–3–3 proteins. FEBS Lett. 435: 110–114.

    Google Scholar 

  • van den Wijngaard, P.W., Bunney, T.D., Roobeek, I., Schonknecht, G. and de Boer, A.H. 2001. Slow vacuolar channels from barley mesophyll cells are regulated by 14–3–3 proteins. FEBS Lett. 488: 100–104.

    Google Scholar 

  • van Heusden, G.P., Griffiths, D.J., Ford, J.C., Chin-A-Woeng, T.F., Schrader, P.A., Carr, A.M. and Steensma, H.Y. 1995. The 14–3–3 proteins encoded by the BMH1 and BMH2 genes are essential in the yeast Saccharomyces cerevisiae and can be replaced by a plant homologue. Eur. J. Biochem. 229: 45–53.

    Google Scholar 

  • Vera-Estrella, R., Barkla, B., Higgins, V.J. and Blumwald, E. 1994. Plant defense response to fungal pathogens: activation of host plasma membrane H+-ATPase by elicitor-induced enzyme dephosphorylation. Plant Physiol. 104: 209–215.

    Google Scholar 

  • Wei, Y.D., Zhang, Z., Andersen, C.H., Schmelzer, E., Gregersen, P.L., Collinge, D.B., Smedegaard-Petersen, V. and Thordal-Christensen, H. 1998. An epidermis/papilla-specific oxalate oxidase-like protein in the defence response of barley attacked by the powdery mildew fungus. Plant Mol. Biol. 36: 101–112

    Google Scholar 

  • Wevelsiep, L., Rüpping, E. and Knogge, W. 1993. Stimulation of barley plasmalemma H+-ATPase by phytotoxic peptides from the fungal pathogen Rhynchosporium secalis. Plant Physiol. 101: 297–301.

    Google Scholar 

  • Xing, T., Higgins, J. and Blumwald, E. 1996. Regulation of plant defence responses to fungal pathogens: two types of protein kinases in the reversible phosphorylation of the host plasma membrane H+-ATPase. Plant Cell 8: 555–564.

    Google Scholar 

  • Xing, T., Higgins, V.J. and Blumwald, E. 1997. Identification of G proteins mediating fungal elicitor-induced dephosphorylation of host plasma membrane H+-ATPase. J. Exp. Bot. 48: 229–237.

    Google Scholar 

  • Zhou, F., Andersen, C.H., Burhenne, K., Fischer, P.H., Collinge, D.B. and Thordal-Christensen, H. 2000. Proton extrusion is an essential signalling component in HR of epidermal single cells in the barley-powdery mildew interaction. Plant J. 23: 245–254.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Finnie, C., Andersen, C.H., Borch, J. et al. Do 14-3-3 proteins and plasma membrane H+-ATPases interact in the barley epidermis in response to the barley powdery mildew fungus?. Plant Mol Biol 49, 137–147 (2002). https://doi.org/10.1023/A:1014938417267

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1014938417267

Navigation