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首页> 外文期刊>Journal of proteome research >Mass spectrometry-based proteomics identifies UPF1 as a critical gene expression regulator in MonoMac 6 cells
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Mass spectrometry-based proteomics identifies UPF1 as a critical gene expression regulator in MonoMac 6 cells

机译:基于质谱的蛋白质组学将UPF1鉴定为MonoMac 6细胞中的关键基因表达调节剂

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摘要

5-Lipoxygenase (5-LO) catalyzes the two initial steps in the biosynthesis of leukotrienes, a group of inflammatory lipid mediators derived from arachidonic acid. Recently, we have demonstrated that 5-LO mRNA expression is regulated by alternative splicing and nonsense-mediated mRNA decay (NMD). In addition to this, 5-LO protein expression was reduced on translational level in UPF1 knockdown cells, suggesting that UPF1 has a positive influence on 5-LO translation. Therefore, a mass spectrometry-based proteomics study was performed to identify compartment-specific protein expression changes upon UPF1 knockdown in differentiated and undifferentiated MM6 cells. The proteomics analysis revealed that the knockdown of UPF1 results in numerous protein changes in the microsomal fraction (~21%) but not in the cytosolic fraction (<1%). The results suggest that UPF1 is a critical gene expression regulator in a compartment-specific way. During differentiation by TGFβ and calcitriol, the majority of UPF1 regulated proteins were adjusted to normal level. This indicates that the translational regulation by UPF1 can potentially be cell differentiation-dependent.
机译:5-脂氧合酶(5-LO)催化白三烯的生物合成中的两个初始步骤,白三烯是一组从花生四烯酸衍生的炎性脂质介质。最近,我们已经证明5-LO mRNA表达是由选择性剪接和无义介导的mRNA衰变(NMD)调节的。除此之外,UPF1敲低细胞中5-LO蛋白的表达降低了翻译水平,这表明UPF1对5-LO翻译具有积极影响。因此,进行了基于质谱的蛋白质组学研究,以鉴定在分化和未分化的MM6细胞中UPF1敲低后,隔室特异性蛋白质表达的变化。蛋白质组学分析表明,敲低UPF1会导致微粒体级分(〜21%)中许多蛋白质变化,而胞质级分(<1%)中无变化。结果表明,UPF1以区室特异性方式是关键的基因表达调节剂。在通过TGFβ和骨化三醇进行分化的过程中,大多数受UPF1调节的蛋白被调节到正常水平。这表明UPF1的翻译调控可能与细胞分化有关。

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