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S-nitrosylation of NADPH oxidase regulates cell death in plant immunity

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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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Figure 1: SNOs positively regulate cell death by hypersensitive response.
Figure 2: Increased SNO concentrations blunt NADPH oxidase activity and reduce ROI accumulation.
Figure 3: S -nitrosylation of AtRBOHD.
Figure 4: The AtRBOHD Cys890Ala mutant shows increased activity during the defence response, amplifying ROI accumulation and cell death development.

Change history

  • 13 October 2011

    The alignment of lane labelling was corrected in Fig. 3c.


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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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Correspondence to Gary J. Loake.

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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).

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