Skip to main content
Log in

Proteomic Analysis of Glutathione S-Transferases of Arabidopsis thaliana Reveals Differential Salicylic Acid-Induced Expression of the Plant-Specific Phi and Tau Classes

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Plant glutathione S-transferases (GSTs) are a large group of multifunctional proteins that are induced by diverse stimuli. Using proteomic approaches we identified 20 GSTs at the protein level in Arabidopsis cell culture with a combination of GST antibody detection, LC-MS/MS analysis of 23–30 kDa proteins and glutathione-affinity chromatography. GSTs identified were from phi, tau, theta, zeta and DHAR sub-sections of the GST superfamily of 53 members. We have uncovered preliminary evidence for post-translational modifications of plant GSTs and show that phosphorylation is unlikely to be responsible. Detailed analysis of GST expression in response to treatment with 0.01–1 mM of the plant defence signal salicylic acid (SA) uncovered some interesting features. Firstly, GSTs appear to display class-specific concentration-dependent SA induction profiles highlighting differences between the large, plant specific phi and tau classes. Secondly, different members of the same class, while sharing similar SA dose responses, may display differences in terms of magnitude and timing of induction, further highlighting the breadth of GST gene regulation. Thirdly, closely related members of the same class (GSTF6 and GSTF7), arising via tandem duplication, may be regulated differently in terms of basal expression levels and also magnitude of induction raising questions about the role of subfunctionalisation within this family. Our results reveal that GSTs exhibit class specific responses to SA treatment suggesting that several mechanisms are acting to induce GSTs upon SA treatment and hinting at class-specific functions for this large and important, yet still relatively elusive gene family.

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

  • Bianchi, M.W., Roux, C. and Vartanian, N. 2002. Drought regulation of GST8, encoding the Arabidopsis homologue of ParC/Nt107 glutathione transferase/peroxidase. Physiol. Plant 116: 96–105.

    Google Scholar 

  • Borsani, O., Valpuesta, V. and Botella, M.A. 2001. Evidence for a role of salicylic acid in the oxidative damage generated by NaCl and osmotic stress in Arabidopsis seedlings. Plant Physiol. 126: 1024–1030.

    Google Scholar 

  • Chen, W. and Singh, K.B. 1999. The auxin, hydrogen peroxide and salicylic acid induced expression of the Arabidopsis GST6 promoter is mediated in part by an ocs element. Plant J. 19: 667–677.

    Google Scholar 

  • Cummins, I., Cole, D.J. and Edwards, R. 1997. Purification of multiple glutathione transferases involved in herbicide detoxification from wheat (Triticum aestivum L.) treated with the safener fenchlorazole-ethyl. Pestic. Biochem. Physiol. 59: 35–49.

    Google Scholar 

  • Dempsey, D., Shah, J. and Klessig, D. 1999. Salicylic acid and disease resistance in plants. Crit. Rev. Plant Sci. 18: 547–575.

    Google Scholar 

  • DeRidder, B.P., Dixon, D.P., Beussman, D.J., Edwards, R. and Goldsbrough, P.B. 2002. Induction of glutathione S-transferases in Arabidopsis by herbicide safeners. Plant Physiol. 130: 1497–1505.

    Google Scholar 

  • Dixon, D.P., Cole, D.J. and Edwards, R. 1998. Purification, regulation and cloning of a glutathione transferase (GST) from maize resembling the auxin-inducible type-III GSTs. Plant Mol. Biol. 36: 75–87.

    Google Scholar 

  • Dixon, D.P., Davis, B.G., Edwards, R. 2002a. Functional divergence in the glutathione transferase superfamily in plants. Identification of two classes with putative functions in redox homeostasis in Arabidopsis thaliana. J. Biol. Chem. 277: 30859–30869.

    Google Scholar 

  • Dixon, D.P., Lapthorn, A. and Edwards, R. 2002b. Plant glutathione transferases. Genome Biol. 3: REVIEWS3004.

  • Edwards, R., Dixon, D.P. and Walbot, V. 2000. Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends Plant Sci. 5: 193–198.

    Google Scholar 

  • Force, A., Lynch, M., Pickett, F.B., Amores, A., Yan, Y.L. and Postlethwait, J. 1999. Preservation of duplicate genes by complementary, degenerative mutations. Genetics 151: 1531–1545.

    Google Scholar 

  • Frova, C. 2003. The plant glutathione transferase gene family: genomic structure, functions, expression and evolution. Physiol. Plant 119: 469–479.

    Google Scholar 

  • Grant, J.J. and Loake, G.J. 2000. Role of reactive oxygen intermediates and cognate redox signaling in disease resistance. Plant Physiol. 124: 21–29.

    Google Scholar 

  • Grant, J.J., Byung-Wook, Y. and Loake, G.J. 2000. Oxidative burst and cognate redox signalling reported by luciferase imaging: identification of a signal network that functions independently of ethylene, SA and Me-JA but is dependent on MAPKK activity. Plant J. 24: 569–582.

    Google Scholar 

  • Kiyosue, T., Yamaguchi-Shinozaki, K. and Shinozaki, K. 1993. Characterization of two cDNAs (ERD11 and ERD13) for dehydration-inducible genes that encode putative glutathione S-transferases in Arabidopsis thaliana L. FEBS Lett. 335: 189–192.

    Google Scholar 

  • Lieberherr, D., Wagner, U., Dubuis, P.-H., Metraux, J.-P. and Mauch, F. 2003. The rapid induction of glutathione S-transferases AtGSTF2 and AtGSTF6 by avirulent Pseudomonas syringae is the result of combined salicylic acid and ethylene signalling. Plant Cell Physiol. 44: 750–757.

    Google Scholar 

  • Loyall, L., Uchida, K., Braun, S., Furuya, M. and Frohnmeyer, H. 2000. Glutathione and a UV light-induced glutathione S-transferase are involved in signaling to chalcone synthase in cell cultures. Plant Cell 12: 1939–1950.

    Google Scholar 

  • Marrs, K.A. 1996. The functions and regulation of glutathione S-transferases in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47: 127–158.

    Google Scholar 

  • Marrs, K.A., Alfenito, M.R., Lloyd, A.M. and Walbot, V. 1995. A glutathione S-transferase involved in vacuolar transfer encoded by the maize gene Bronze-2. Nature 375: 397–400.

    Google Scholar 

  • McGonigle, B., Keeler, S.J., Cindy Lau, S.-M., Koeppe, M.K. and O'Keefe, D.P. 2000. A genomics approach to the comprehensive analysis of the glutathione S-transferase gene family in soybean and maize. Plant Physiol. 124: 1105–1120.

    Google Scholar 

  • Pfaffl, M.W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucl. Acids Res. 29: e45.

    Google Scholar 

  • Pfaffl, M.W., Horgan, G.W. and Dempfle, L. 2002. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucl. Acids Res. 30: e36.

    Google Scholar 

  • Reuber, T.L., Plotnikova, J.M., Dewdney, J., Rogers, E.E., Wood, W. and Ausubel, F.M. 1998. Correlation of defense gene induction defects with powdery mildew susceptibility in Arabidopsis enhanced disease susceptibility mutants. Plant J. 16: 473–485.

    Google Scholar 

  • Schulenberg, B., Aggeler, R., Beechem, J.M., Capaldi, R.A. and Patton, W.F. 2003. Analysis of steady-state protein phosphorylation in mitochondria using a novel fluorescent phosphosensor dye. J. Biol. Chem. 278: 27251–27255.

    Google Scholar 

  • Senaratna, T., Touchell, D., Bunn, E. and Dixon, K. 2000. Acetyl salicylic acid (Aspirin) and salicylic acid induce multiple stress tolerance in bean and tomato plants. Plant Growth Regul. 30: 157–161.

    Google Scholar 

  • Sharma, Y.K., Leon, J., Raskin, I. and Davis, K.R. 1996. Ozone-induced responses in Arabidopsis thaliana: the role of salicylic acid in the accumulation of defense-related transcripts and induced resistance. Proc. Natl. Acad. Sci. USA 93: 5099–5104.

    Google Scholar 

  • Sheehan, D., Meade, G., Foley, V.M. and Dowd, C.A. 2001. Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem. J. 360: 1–16.

    Google Scholar 

  • Smith, A.P., Nourizadeh, S.D., Peer, W.A., Xu, J., Bandyopadhyay, A., Murphy, A.S. and Goldsbrough, P.B. 2003. Arabidopsis AtGSTF2 is regulated by ethylene and auxin, and encodes a glutathione S-transferase that interacts with flavonoids. Plant J. 36: 433–442.

    Google Scholar 

  • Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F. and Higgins, D.G. 1997. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl. Acids Res. 24: 4876–4882.

    Google Scholar 

  • Wagner, U., Edwards, R., Dixon, D.P. and Mauch, F. 2002. Probing the diversity of the Arabidopsis glutathione S-transferase gene family. Plant Mol. Biol. 49: 515–532.

    Google Scholar 

  • Wessel, D. and Flugge, U.I. 1984. A Method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal. Biochem. 138: 141–143.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to A. Harvey Millar.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sappl, P.G., Oñate-Sánchez, L., Singh, K.B. et al. Proteomic Analysis of Glutathione S-Transferases of Arabidopsis thaliana Reveals Differential Salicylic Acid-Induced Expression of the Plant-Specific Phi and Tau Classes. Plant Mol Biol 54, 205–219 (2004). https://doi.org/10.1023/B:PLAN.0000028786.57439.b3

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:PLAN.0000028786.57439.b3

Navigation