Abstract
Changes in redox status are a conspicuous feature of immune responses in a variety of eukaryotes1,2, but the associated signalling mechanisms are not well understood. In plants, attempted microbial infection triggers the rapid synthesis of nitric oxide3,4 and a parallel accumulation of reactive oxygen intermediates, the latter generated by NADPH oxidases related to those responsible for the pathogen-activated respiratory burst in phagocytes5. Both nitric oxide and reactive oxygen intermediates have been implicated in controlling the hypersensitive response, a programmed execution of plant cells at sites of attempted infection3,5,6. However, the molecular mechanisms that underpin their function and coordinate their synthesis are unknown. Here we show genetic evidence that increases in cysteine thiols modified using nitric oxide, termed S-nitrosothiols, facilitate the hypersensitive response in the absence of the cell death agonist salicylic acid and the synthesis of reactive oxygen intermediates. Surprisingly, when concentrations of S-nitrosothiols were high, nitric oxide function also governed a negative feedback loop limiting the hypersensitive response, mediated by S-nitrosylation of the NADPH oxidase, AtRBOHD, at Cys 890, abolishing its ability to synthesize reactive oxygen intermediates. Accordingly, mutation of Cys 890 compromised S-nitrosothiol-mediated control of AtRBOHD activity, perturbing the magnitude of cell death development. This cysteine is evolutionarily conserved and specifically S-nitrosylated in both human and fly NADPH oxidase, suggesting that this mechanism may govern immune responses in both plants and animals.
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Change history
13 October 2011
The alignment of lane labelling was corrected in Fig. 3c.
References
MacMicking, J. D. et al. Identification of nitric oxide synthase as a protective locus against tuberculosis. Proc. Natl Acad. Sci. USA 94, 5243–5248 (1997)
Tada, Y. et al. Plant immunity requires conformational changes of NPR1 via S-nitrosylation and thioredoxins. Science 321, 952–956 (2008)
Delledonne, M. Xia, Y. Dixon, R. A. & Lamb, C. J. Nitric oxide functions as signal in plant disease resistance. Nature 394, 585–588 (1998)
Durner, J., Wendehenne, D. & Klessig, D. F. Defense gene induction in tobacco by nitric oxide, cyclic GMP and cyclic ADP ribose. Proc. Natl Acad. Sci. USA 95, 10328–10333 (1998)
Torres, M. A., Dangl, J. L. & Jones, J. D. Arabidopsis gp91phox homologues AtrbohD and AtrbohF are required for accumulation of reactive oxygen intermediates in the plant defense response. Proc. Natl Acad. Sci. USA 99, 517–522 (2002)
Delledonne, M., Zeier, J., Marocco, A. & Lamb, C. Signal interactions between nitric oxide and reactive oxygen intermediates in the plant hypersensitive disease resistance response. Proc. Natl Acad. Sci. USA 98, 13454–13459 (2001)
Leitner, M., Vandelle, E., Gaupels, F., Bellin, D. & Delledonne, M. NO signals in the haze: nitric oxide signalling in plant defence. Curr. Opin. Plant Biol. 12, 451–458 (2009)
Feechan, A. et al. A central role for S-nitrosothiols in plant disease resistance. Proc. Natl Acad. Sci. USA 102, 8054–8059 (2005)
He, Y. et al. Nitric oxide represses the Arabidopsis floral transition. Science 305, 1968–1971 (2004)
Grant, M. R. et al. Structure of the Arabidopsis RPM1 gene enabling dual specificity disease resistance. Science 269, 843–846 (1995)
Gassmann, W., Hinsch, M. E. & Staskawicz, B. J. The Arabidopsis RPS4 bacterial-resistance gene is a member of the TIR-NBS-LRR family of disease resistance genes. Plant J. 20, 265–277 (1999)
Liu, L. et al. Essential roles of S-nitrosothiols in vascular homeostasis and endotoxic shock. Cell 116, 617–628 (2004)
Shirasu, K., Nakajima, H., Rajasekhar, V. K., Dixon, R. A. & Lamb, C. J. Salicylic acid potentiates an agonist-dependent gain control that amplifies pathogen signals in the activation of defense mechanisms. Plant Cell 9, 261–270 (1997)
Wildermuth, M. C., Dewdney, J., Wu, G. & Ausubel, F. M. Isochorismate synthase is required to synthesize salicylic acid for plant defence. Nature 414, 562–565 (2001)
Yu, I. C., Parker, J. & Bent, A. F. Gene-for-gene disease resistance without the hypersensitive response in Arabidopsis dnd1 mutant. Proc. Natl Acad. Sci. USA 95, 7819–7824 (1998)
Holub, E. B. Beynon, J. L. & Crute, I. R. Phenotypic and genotypic characterization of interactions between isolates of Peronospora parasitica and accessions of Arabidopsis thaliana . Mol. Plant Microbe Interact. 7, 223–239 (1994)
Keller, H. et al. Pathogen-induced elicitin production in transgenic tobacco generates a hypersensitive response and nonspecific disease resistance. Plant Cell 11, 223–236 (1999)
Nawrath, C. & Metraux, J. P. Salicylic acid induction-deficient mutants of Arabidopsis express PR-2 and PR-5 and accumulate high levels of camalexin after pathogen inoculation. Plant Cell 11, 1393–1404 (1999)
Wang, Y.-Q. et al. S-nitrosylation of AtSABP3 antagonises the expression of plant immunity. J. Biol. Chem. 284, 2131–2137 (2009)
Romero-Puertas, M. C. et al. S-nitrosylation of peroxiredoxin II E promotes peroxynitrite-mediated tyrosine nitration. Plant Cell 19, 4120–4130 (2007)
Lindermayr, C., Sell, S., Müller, B., Leister, D. & Durner, J. Redox regulation of the NPR1–TGA1 system of Arabidopsis thaliana by nitric oxide. Plant Cell 22, 2894–2907 (2010)
Jaffrey, S. R., Erdjument-Bromge, H., Ferris, C. D., Tempst, P. & Snyder, S. H. Protein S-nitrosylation: a physiological signal for neuronal nitric oxide. Nature Cell Biol. 3, 193–197 (2001)
Selemidis, S., Dusting, G. J., Peshavariya, H., Kemp-Harper, B. K. & Drummond, G. R. Nitric oxide suppresses NADPH oxidase-dependent superoxide production by S-nitrosylation in human endothelial cells. Cardiovasc. Res. 75, 349–358 (2007)
Ingelman, M., Bianchi, V. & Eklund, H. The three-dimensional structure of flavodoxin reductase from Escherichia coli at 1.7 Å resolution. J. Mol. Biol. 268, 147–157 (1997)
Zhen, L., Yu, L. & Dinauer, M. C. Probing the role of the carboxyl terminus of the gp91 phox subunit of neutrophil flavocytochrome b 558 using site-directed mutagenesis. J. Biol. Chem. 273, 6575–6581 (1998)
Matthews, J. R. et al. Inhibition of NF-κβ DNA binding by nitric oxide. Nucleic Acids Res. 24, 2236–2242 (1996)
Mannick, J. B. et al. Fas-induced caspase denitrosylation. Science 284, 651–654 (1999)
Yun, B.-W. et al. Loss of actin cytoskeletal function and EDS1 activity, in combination, severely compromises non-host resistance in Arabidopsis against wheat powdery mildew. Plant J. 34, 768–777 (2003)
Aboul-Soud, M. A. M., Cook, K. & Loake, G. J. Measurement of salicylic acid by a high-performance liquid chromatography procedure based on ion-exchange. Chromatographia 59, 129–133 (2004)
Liu, L. et al. Essential roles of S-nitrosothiols in vascular homeostasis and endotoxic shock. Cell 116, 617–628 (2004)
Foissner, I., Wendehenne, D., Langebartels, C. & Durner, J. In vivo imaging of an elicitor-induced nitric oxide burst in tobacco. Plant J. 23, 817–824 (2000)
Whalen, M. C., Innes, R. W., Bent, A. F. & Staskawicz, B. J. Identification of Pseudomonas syringae pathogens of Arabidopsis and a bacterial locus determining avirulence on both Arabidopsis and soybean. Plant Cell 3, 49–59 (1991)
Dellagi, A., Brisset, M.-N., Jean-Pierre Paulin, J.-P. & Expert, D. Dual role of desferrioxamine in Erwinia amylovora pathogenicity. Mol. Plant Microbe Interact. 11, 734–742 (1998)
Liu, Q., Li, M., Leibham, D., Cortez, D. & Elledge, S. The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without restriction enzymes. Curr. Biol. 8, 1300–1309 (1998)
Sagi, M. & Fluhr, R. Superoxide production by plant homologues of the gp91phox NADPH oxidase. Modulation of activity by calcium and by tobacco mosaic virus infection. Plant Physiol. 126, 1281–1290 (2001)
Chen, Y. Y., Huang, Y. F., Khoo, K. H. & Meng, T. C. Mass spectrometry-based analyses for identifying and characterizing S-nitrosylation of protein tyrosine phosphatases. Methods 42, 243–249 (2007)
Shen, A. L. &. Kasper, C. B. Differential contribution of NADPH-cytochrome P450 oxidoreductase FAD binding site residues to flavin binding and catalysis. J. Biol. Chem. 275, 41087–41091 (2000)
Kelley, L. A. & Sternberg, M. J. Protein structure prediction on the Web: a case study using the Phyre server. Nature Protocols 4, 363–371 (2009)
Guex, N. & Peitsch, M. C. SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 18, 2714–2723 (1997)
Cole, C., Barber, J. D. & Barton, G. J. The Jpred 3 secondary structure prediction server. Nucleic Acids Res. 36, W197–W201 (2008)
Acknowledgements
We would like to acknowledge R. Innes and W. Gassmann for Pst DC3000 strains expressing either avrB or avrRps4, respectively. Arabidopsis transfer DNA insertion mutants were obtained from SAIL (Syngeneta) populations. We thank M. Tör for the H. arabidopsidis isolate Emwa1, and K. Kanchanawatee for the Drosophila cDNA clone and associated mutant. A.F. was supported by a Biotechnology and Biological Sciences Research Council (BBSRC) CASE studentship. B.-W.Y. and E.K. were funded by BBSRC grant BB/D011809/1 awarded to G.J.L. J.W.M. received a grant from the Physical Sciences Research Council (EPSRC). M. Yu was the recipient of a Darwin Trust Scholarship. T.L.B. was supported by BBSRC and EPSRC grant BB/D019621/1. N.B.B.S. and M. Yin were funded by a Ministry of Education Malaysia scholarship and a Torrance Scholarship, respectively.
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G.J.L. designed the research and wrote the paper, and with B.-W.Y. planned experiments and analyses. B.-W.Y. conducted the majority of experiments. A.F., M. Yin, N.B.B.S., J.-G.K., J.W.M., M. Yu, E.K. and T.L.B. conducted experiments. S.H.S. generated and interrogated three-dimensional models. J.A.P. was the industrial supervisor of A.F. All authors, especially B.-W.Y. and S.S., commented on the manuscript.
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Yun, BW., Feechan, A., Yin, M. et al. S-nitrosylation of NADPH oxidase regulates cell death in plant immunity. Nature 478, 264–268 (2011). https://doi.org/10.1038/nature10427
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DOI: https://doi.org/10.1038/nature10427
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