Activation of mitogen-activated protein kinase by H2O2 - Role in cell survival following oxidant injury
Activation of mitogen-activated protein kinase by H2O2 - Role in cell survival following oxidant injury
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DOI：
10.1074/jbc.271.7.3604
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发表时间：
1996-02-23
影响因子：
4.8
通讯作者：
Holbrook, NJ
中科院分区：
文献类型：
--
作者：
Guyton, KZ;Liu, YS;Holbrook, NJ
丝裂原活化蛋白激酶（MAPK）家族由控制细胞对增殖和应激信号的反应的关键调节蛋白组成。在这项研究中，我们调查了控制MAPK激活的因素，通过H2 O2和探讨的影响，改变激酶激活的途径对细胞存活后H2 O2暴露。有效激活细胞外信号调节蛋白激酶（ERK 2）在H2 O2处理10分钟内发生（10-20倍），随后迅速失活，H2 O2在几种细胞类型中激活ERK 2，另外，H_2O_2使MAPK依赖基因c-jun、c-fos、和MAPK磷酸酶-1。苏拉明预处理完全抑制H2 O2刺激ERK 2，突出了生长因子受体在这种激活中的作用，此外，ERK 2激活H2 O2被阻断预处理与N-乙酰半胱氨酸，邻菲咯啉，或甘露醇，表明金属催化的自由基形成介导的H2 O2的信号转导的启动。在PC 12细胞中，H2 O2刺激的ERK 2激活被显性负性Ras-N-17等位基因的诱导型或组成型表达所消除。有趣的是，PC 12/Ras-N-17细胞比野生型PC 12细胞对H2 O2毒性更敏感。此外，NIH 3 T3细胞表达组成型活性MAPK激酶（MEK，ERK的直接上游调节）更耐H2 O2毒性，而那些表达激酶缺陷的MEK更敏感，比表达野生型MEK的细胞。两者合计，这些研究提供了深入了解MAPK的调节机制H2 O2，并建议ERK在氧化损伤后的细胞存活中起着至关重要的作用。
The mitogen-activated protein kinase (MAPK) family is comprised of key regulatory proteins that control the cellular response to both proliferation and stress signals. In this study we investigated the factors controlling MAPK activation by H2O2 and explored the impact of altering the pathways to kinase activation on cell survival following H2O2 exposure. Potent activation (10-20-fold) of extracellular signal-regulated protein kinase (ERK2) occurred within 10 min of H2O2 treatment, whereupon rapid inactivation ensued, H2O2 activated ERK2 in several cell types and also moderately activated (3-5-fold) both c-Jun N-terminal kinase and p38/RK/CSBP, Additionally, H2O2 increased the mRNA expression of MAPK-dependent genes c-jun, c-fos, and MAPK phosphatase-1. Suramin pretreatment completely inhibited H2O2 stimulation of ERK2, highlighting a role for growth factor receptors in this activation, Further, ERK2 activation by H2O2 was blocked by pretreatment with either N-acetyl-cysteine, o-phenanthroline, or mannitol, indicating that metal-catalyzed free radical formation mediates the initiation of signal transduction by H2O2. H2O2-stimulated activation of ERK2 was abolished in PC12 cells by inducible or constitutive expression of the dominant negative Ras-N-17 allele. Interestingly, PC12/Ras-N-17 cells were more sensitive than wild-type PC12 cells to H2O2 toxicity. Moreover, NIH 3T3 cells expressing constitutively active MAPK kinase (MEK, the immediate upstream regulator of ERK) were more resistant to H2O2 toxicity, while those expressing kinase-defective MEK were more sensitive, than cells expressing wild-type MEK. Taken together, these studies provide insight into mechanisms of MAPK regulation by H2O2 and suggest that ERK plays a critical role in cell survival following oxidant injury.