Post-transcriptional regulation of Transforming Growth Factor Beta-1 by microRNA-744.
Post-transcriptional regulation of Transforming Growth Factor Beta-1 by microRNA-744.
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DOI：
10.1371/journal.pone.0025044
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Fraser DJ
中科院分区：
文献类型：
--
作者：
Martin J;Jenkins RH;Bennagi R;Krupa A;Phillips AO;Bowen T;Fraser DJ
转化生长因子-β-1是一种多效性细胞因子，在伤口愈合、炎症以及包括癌症和进行性组织纤维化在内的关键病理过程中发挥重要作用。转化生长因子-β-1是转录后调控的，但其潜在的机制仍不完全清楚。在此之前，我们已经广泛地描述了转化生长因子-β1在肾脏中合成的转录后调控，有证据表明在糖尿病肾病的背景下，近端肾小管上皮细胞的翻译抑制得到缓解。在本研究中，我们研究了转化生长因子-β13‘非编码区(3’非编码区)在肿瘤发生发展中的作用。已经报道了转化生长因子-β1的两个不同的3‘非编码区长度，分别为543和137个核苷酸。绝对定量研究表明，虽然在不同的人类细胞类型和广泛的组织中都可以检测到这两种UTR长度，但短型在肾脏和其他地方占主导地位。经转化生长因子-β-1自身诱导后，这两种形式的表达均上调，但短/长UTR值保持不变。将短非编码区整合到荧光素酶报告载体中，显著减少了报告蛋白的合成，而对RNA含量没有显著影响，这表明转录后抑制。在电子计算机方法中，发现了位于转化生长因子-β1 3‘非编码区近端的miR-744的多个结合位点。从人体组织的RNA中筛选出广泛表达的miR-744。MiR-744可抑制内源性转化生长因子-β-1的合成，而直接靶向转化生长因子-β1的实验表明，miR-744可降低转化生长因子-β1 3‘非编码区报告基因的活性。这项工作确定了miR-744指导的转化生长因子-β1的转录后调控，鉴于转化生长因子-β1的细胞反应的多效性，这一调控具有广泛的潜在意义。
Transforming Growth Factor Beta-1 (TGF-β1) is a pleiotropic cytokine that is of central importance in wound healing, inflammation, and in key pathological processes including cancer and progressive tissue fibrosis. TGF-β1 is post-transcriptionally regulated, but the underlying mechanisms remain incompletely defined. Previously, we have extensively delineated post-transcriptional regulation of TGF-β1 synthesis in the kidney, with evidence for relief of translational repression in proximal tubular cells in the context of diabetic nephropathy. In this study, we have investigated the role of the TGF-β1 3′Untranslated Region (3′UTR). Two different 3′UTR lengths have been reported for TGF-β1, of 543 and 137 nucleotides. Absolute quantification showed that, while both UTR lengths were detectable in various human cell types and in a broad range of tissues, the short form predominated in the kidney and elsewhere. Expression of both forms was up-regulated following auto-induction by TGF-β1, but the short:long UTR ratio remained constant. Incorporation of the short UTR into a luciferase reporter vector significantly reduced reporter protein synthesis without major effect on RNA amount, suggesting post-transcriptional inhibition. In silico approaches identified multiple binding sites for miR-744 located in the proximal TGF-β1 3′UTR. A screen in RNA from human tissues showed widespread miR-744 expression. miR-744 transfection inhibited endogenous TGF-β1 synthesis, while direct targeting of TGF-β1 was shown in separate experiments, in which miR-744 decreased TGF-β1 3′UTR reporter activity. This work identifies miR-744-directed post-transcriptional regulation of TGF-β1 which, given the pleiotropic nature of cellular responses to TGF-β1, is potentially widely significant.
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