State Key Laboratory of Plant Genomics and National Plant Gene Research Center (Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Cell Res. 2009 Dec;19(12):1377-87. doi: 10.1038/cr.2009.117. Epub 2009 Oct 6.
Metabolism of S-nitrosoglutathione (GSNO), a major biologically active nitric oxide (NO) species, is catalyzed by the evolutionally conserved GSNO reductase (GSNOR). Previous studies showed that the Arabidopsis GSNOR1/HOT5 gene regulates salicylic acid signaling and thermotolerance by modulating the intracellular S-nitrosothiol level. Here, we report the characterization of the Arabidopsis paraquat resistant2-1 (par2-1) mutant that shows an anti-cell death phenotype. The production of superoxide in par2-1 is comparable to that of wild-type plants when treated by paraquat (1,1'-dimethyl-4,4'-bipyridinium dichloride), suggesting that PAR2 acts downstream of superoxide to regulate cell death. PAR2, identified by positional cloning, is shown to be identical to GSNOR1/HOT5. The par2-1 mutant carries a missense mutation in a highly conserved glycine, which renders the mutant protein unstable. Compared to wild type, par2-1 mutant has a higher NO level, as revealed by staining with 4,5-diaminofluorescein diacetate. Consistent with this result, wild-type plants treated with an NO donor display resistance to paraquat. Interestingly, the GSNOR1/HOT5/PAR2 protein level, other than its steady-state mRNA level, is induced by paraquat, but is reduced by NO donors. Taken together, these results suggest that GSNOR1/HOT5/PAR2 plays an important role in regulating cell death in plant cells through modulating intracellular NO level.
S-亚硝基谷胱甘肽(GSNO)是一种主要的生物活性一氧化氮(NO)物质,其代谢由进化保守的 GSNO 还原酶(GSNOR)催化。先前的研究表明,拟南芥 GSNOR1/HOT5 基因通过调节细胞内 S-亚硝基硫醇水平来调节水杨酸信号和耐热性。在这里,我们报道了拟南芥百草枯抗性 2-1(par2-1)突变体的特征,该突变体表现出抗细胞死亡表型。当用百草枯(1,1'-二甲基-4,4'-联吡啶二氯化物)处理时,par2-1 中的超氧阴离子的产生与野生型植物相当,这表明 PAR2 作用于超氧阴离子下游以调节细胞死亡。通过定位克隆鉴定的 PAR2 与 GSNOR1/HOT5 相同。par2-1 突变体携带高度保守的甘氨酸中的错义突变,使突变蛋白不稳定。与野生型相比,par2-1 突变体的 NO 水平更高,这是通过用 4,5-二氨基荧光素二乙酸酯染色揭示的。与该结果一致,用 NO 供体处理的野生型植物对百草枯表现出抗性。有趣的是,除了其稳定状态的 mRNA 水平外,百草枯诱导 GSNOR1/HOT5/PAR2 蛋白水平升高,但被 NO 供体降低。总之,这些结果表明,GSNOR1/HOT5/PAR2 通过调节细胞内 NO 水平在调节植物细胞死亡中起重要作用。