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Regulation of RNA polymerase III transcription during transformation of human IMR90 fibroblasts with defined genetic elements

机译:用定义遗传元素的人IMR90成纤维细胞转化中RNA聚合酶III转录的调节

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摘要

RNA polymerase (Pol) III transcribes small untranslated RNAs that are essential for cellular homeostasis and growth. Its activity is regulated by inactivation of tumor suppressor proteins and overexpression of the oncogene c-MYC, but the concerted action of these tumor-promoting factors on Pol III transcription has not yet been assessed. In order to comprehensively analyse the regulation of Pol III transcription during tumorigenesis we employ a model system that relies on the expression of five genetic elements to achieve cellular transformation. Expression of these elements in six distinct transformation intermediate cell lines leads to the inactivation of TP53, RB1, and protein phosphatase 2A, as well as the activation of RAS and the protection of telomeres by TERT, thereby conducting to full tumoral transformation of IMR90 fibroblasts. Transformation is accompanied by moderately enhanced levels of a subset of Pol III-transcribed RNAs (7SK; MRP; H1). In addition, mRNA and/or protein levels of several Pol III subunits and transcription factors are upregulated, including increased protein levels of TFIIIB and TFIIIC subunits, of SNAPC1 and of Pol III subunits. Strikingly, the expression of POLR3G and of SNAPC1 is strongly enhanced during transformation in this cellular transformation model. Collectively, our data indicate that increased expression of several components of the Pol III transcription system accompanied by a 2-fold increase in steady state levels of a subset of Pol III RNAs is sufficient for sustaining tumor formation.
机译:RNA聚合酶(Pol)III转录对细胞内环境稳定和生长至关重要的小的未翻译RNA。其活性受抑癌蛋白失活和癌基因c-MYC过度表达的调节,但这些促瘤因子对Pol III转录的协同作用尚未评估。为了全面分析Pol III在肿瘤发生过程中的转录调控,我们采用了一个模型系统,该系统依赖于五种遗传元素的表达来实现细胞转化。这些元件在六种不同的转化中间细胞系中的表达导致TP53、RB1和蛋白磷酸酶2A失活,以及RAS的激活和TERT对端粒的保护,从而实现IMR90成纤维细胞的完全肿瘤转化。转化伴随着Pol III转录RNA亚群(7SK;MRP;H1)水平的适度增强。此外,几个Pol III亚单位和转录因子的mRNA和/或蛋白质水平上调,包括TFIIIB和TFIIIC亚单位、SNAPC1和Pol III亚单位的蛋白质水平升高。引人注目的是,在这种细胞转化模型中,POLR3G和SNAPC1的表达在转化过程中显著增强。总的来说,我们的数据表明,Pol III转录系统的几个组成部分的表达增加,伴随着Pol III RNA亚群的稳态水平增加2倍,足以维持肿瘤的形成。

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