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Post-transcriptional regulation of HSP70 expression following oxidative stress in SH-SY5Y cells: the potential involvement of the RNA-binding protein HuR.

机译:SH-SY5Y细胞氧化应激后HSP70表达的后转录调节:RNA结合蛋白患者的潜在累及。

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Brain aging is associated with a progressive imbalance between intracellular concentration of Reactive Oxygen Species (ROS) and cells ability to activate defensive genes. Heat Shock Protein 70 (HSP70) has been shown to act as a fundamental defensive mechanism for neurons exposed to an oxidant challenge, and its expression decreases during senescence. In the present report we show that the RNA-binding protein ELAV/HuR can affect, post-transcriptionally, the fate of HSP70 mRNA following H(2)O(2)-mediated oxidative stress in SH-SY5Y human neuroblastoma cells. As a consequence of H(2)O(2) treatment (1mM for 30 minutes), HSP70 mRNA accumulates in the ribosomes associated to the cytoskeleton, where parallel Western blotting experiments reveal statistically significant increase for both HuR and HSP70 protein levels. We also confirm the capability of HuR to bind to HSP70 mRNA, and describe how the biological effect of this ELAV protein on the HSP70 mRNA could be due to a direct phosphorylation in serine/threonine residues of HuR itself by the early (10 minutes) H(2)O(2)-mediated activation of PKC alpha. Our findings shed light on the post-transcriptional regulation of HSP70 expression, suggesting the existence of a new molecular cascade -involving PKC/HuR/HSP70- that possibly represents an early event in the cellular response to H(2)O(2)-mediated oxidative stress in SH-SY5Y human neuroblastoma cells. The present results lead us to speculate that an impairment in this regulatory mechanism might directly contribute to the defective cellular response to oxidative stress, thus helping to dissect a potential tool useful to counteract some aspects associated to cerebral senescence.
机译:脑衰老与细胞内浓度(ROS)和激活防守基因的能力之间的逐渐失衡相关。已经证明了热休克蛋白70(HSP70)作为暴露于氧化剂攻击的神经元的基本防御机制,其表达在衰老期间减少。在本报告中,我们表明RNA结合蛋白ELAV / HUR可以在转录后,HSP70 mRNA的命运在SH-SY5Y人神经母细胞瘤细胞中的H(2)O(2)介导的氧化应激之后。由于H(2)O(2)处理(1mm持续30分钟),Hsp70 mRNA在与细胞骨架相关的核糖体中累积,并且平行的Western印迹实验显示Hur和Hsp70蛋白质水平的统计上显着增加。我们还证实了HUR与HSP70 mRNA结合的能力,并描述了该ELAV蛋白在HSP70 mRNA上的生物学效应如何是由于在早期(10分钟)H受尿素/苏氨酸残留物中的直接磷酸化(2)O(2)介导的PKCα的激活。我们的研究结果揭示了HSP70表达的转录后调节,表明存在新的分子级联 - 植物PKC / HUS / HSP70-这可能代表H(2)O(2)的细胞反应中的早期事件 - 介导的SH-SY5Y人神经母细胞瘤细胞中的氧化应激。目前的结果引导我们推测该调节机制中的损伤可能直接有助于对氧化应激的缺陷的细胞反应有助于解剖可用于抵消与脑衰老相关的一些方面的潜在工具。

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