NO-mediated apoptosis in yeast
NO-mediated apoptosis in yeast
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DOI：
10.1242/jcs.010926
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发表时间：
2007-09-15
影响因子：
4
通讯作者：
Ludovico, Paula
中科院分区：
文献类型：
--
作者：
Almeida, Bruno;Buettner, Sabrina;Ludovico, Paula
一氧化氮（NO）是一种小分子，在生物系统的多种生理功能中具有独特的作用，其中包括细胞凋亡信号级联的控制。通过结合蛋白质组学、遗传学和生物化学方法，我们证明了NO和甘油醛-3-磷酸脱氢酶（GAPDH）是酵母细胞凋亡的重要介质。使用间接的方法和NO选择性电极，我们目前的结果表明，过氧化氢诱导的凋亡细胞合成NO是相关的一氧化氮合酶（NOS）样的活性，通过使用一个经典的NOS试剂盒检测证明。此外，我们的研究结果表明，酵母GAPDH是一个目标的广泛的蛋白水解后H2 O2诱导的细胞凋亡，并进行S-亚硝化。用N-ω-硝基-L-精氨酸甲酯阻断NO合成导致GAPDH S-亚硝化和细胞内活性氧（ROS）积累减少，从而增加存活。这些结果表明，NO信号和GAPDH S-亚硝化与H2 O2诱导的凋亡细胞死亡。有证据表明，NO和GAPDH S-亚硝化也介导的细胞死亡，在时间寿命指向一个生理作用的NO在酵母细胞凋亡。
Nitric oxide (NO) is a small molecule with distinct roles in diverse physiological functions in biological systems, among them the control of the apoptotic signalling cascade. By combining proteomic, genetic and biochemical approaches we demonstrate that NO and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are crucial mediators of yeast apoptosis. Using indirect methodologies and a NO-selective electrode, we present results showing that H2O2-induced apoptotic cells synthesize NO that is associated to a nitric oxide synthase (NOS)-like activity as demonstrated by the use of a classical NOS kit assay. Additionally, our results show that yeast GAPDH is a target of extensive proteolysis upon H2O2-induced apoptosis and undergoes S-nitrosation. Blockage of NO synthesis with N-omega-nitro-L-arginine methyl ester leads to a decrease of GAPDH S-nitrosation and of intracellular reactive oxygen species (ROS) accumulation, increasing survival. These results indicate that NO signalling and GAPDH S-nitrosation are linked with H2O2-induced apoptotic cell death. Evidence is presented showing that NO and GAPDH S-nitrosation also mediate cell death during chronological life span pointing to a physiological role of NO in yeast apoptosis.