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首页> 外文期刊>Molecular and Cellular Biology >Posttranscriptional regulation of urokinase receptor mRNA: identification of a novel urokinase receptor mRNA binding protein in human mesothelioma cells.
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Posttranscriptional regulation of urokinase receptor mRNA: identification of a novel urokinase receptor mRNA binding protein in human mesothelioma cells.

机译：尿激酶受体mRNA的转录后调控：人间皮瘤细胞中新型尿激酶受体mRNA结合蛋白的鉴定。

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Treatment of human pleural mesothelioma (MS-1) cells with phorbol myristate acetate (PMA) and cycloheximide results in 17- and 10-fold, respectively, increases in steady-state expression of urokinase-type plasminogen activator receptor (uPAR) mRNA. Studies of transcriptional inhibition by actinomycin D showed four- and sixfold extensions of uPAR mRNA half-life in MS-1 cells treated with PMA and cycloheximide, respectively, suggesting that uPAR gene expression involves a posttranscriptional regulatory mechanism. Using gel mobility shift and UV cross-linking assays, we identified a 50-kDa uPAR mRNA binding protein (uPAR mRNABp) that selectively bound to a 51-nucleotide (nt) fragment of mRNA corresponding to the uPAR coding region. We investigated the possibility that this 51-nt protein binding fragment of uPAR mRNA contains regulatory information for message stability. Chimeric beta-globin/uPAR/beta-globin mRNA containing the 51-nt protein binding fragment was able to destabilize otherwise stable beta-globin mRNA. Conversely, a control chimeric beta-globin/uPAR/beta-globin mRNA containing a 51-nt fragment of the uPAR coding region that does not bind uPAR mRNABp was stable under identical conditions. Binding of uPAR mRNABp to uPAR mRNA was abolished after treatment with cycloheximide and rapidly down-regulated by PMA. These data suggest that the 51-nt protein binding fragment of uPAR mRNA may be involved in mRNA turnover as well as in cycloheximide-induced uPAR message stabilization. Our results indicate a novel mechanism of uPAR gene regulation in which cis elements within a 51-nt coding region interact with a uPAR mRNABp to regulate uPAR message stability.
机译：用佛波肉豆蔻酸酯乙酸盐（PMA）和环己酰亚胺对人胸膜间皮瘤（MS-1）细胞的治疗分别使尿激酶型纤溶酶原激活剂受体（uPAR）mRNA的稳态表达增加17倍和10倍。放线菌素D抑制转录的研究表明，分别用PMA和环己酰亚胺处理的MS-1细胞中uPAR mRNA半衰期延长了4倍和6倍，这表明uPAR基因表达涉及转录后调控机制。使用凝胶迁移率变化和UV交联测定，我们确定了一个50 kDa的uPAR mRNA结合蛋白（uPAR mRNABp），该蛋白选择性结合与uPAR编码区相对应的mRNA的51个核苷酸（nt）片段。我们调查了uPAR mRNA的这一51 nt蛋白结合片段包含信息稳定性的调节信息的可能性。包含51-nt蛋白结合片段的嵌合β-珠蛋白/ uPAR /β-珠蛋白mRNA能够破坏稳定的β-珠蛋白mRNA的稳定性。相反，包含不结合uPAR mRNABp的uPAR编码区51-nt片段的对照嵌合β-珠蛋白/ uPAR /β-珠蛋白mRNA在相同条件下是稳定的。用环己酰亚胺处理后，uPAR mRNABp与uPAR mRNA的结合被取消，并被PMA快速下调。这些数据表明，uPAR mRNA的51-nt蛋白结合片段可能参与mRNA转换以及环己酰亚胺诱导的uPAR信息稳定。我们的研究结果表明uPAR基因调控的一种新机制，其中51 nt编码区域内的顺式元素与uPAR mRNABp相互作用以调控uPAR消息的稳定性。

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