Characterization of the 5′-untranslated region of YB-1 mRNA and autoregulation of translation by YB-1 protein
Characterization of the 5′-untranslated region of YB-1 mRNA and autoregulation of translation by YB-1 protein
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DOI：
10.1093/nar/gkh223
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发表时间：
2004-01-01
影响因子：
14.9
通讯作者：
Uchiumi, T
中科院分区：
文献类型：
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作者：
Fukuda, T;Ashizuka, M;Uchiumi, T
真核生物Y盒结合蛋白YB-1参与多种生物学过程，包括DNA修复、细胞增殖以及转录和翻译的调节。YB-1蛋白在人类细胞中广泛表达，在细胞增殖和转化中起作用。它的浓度被认为是在转录和翻译水平高度调节。因此，我们研究了YB-1 mRNA的5 '-UTR是否影响YB-1蛋白的翻译，从而影响表达水平。将连接至YB-1 mRNA 5 '-UTR的荧光素酶mRNA用作报告构建体。全长YB-1 5 '-UTR（331个碱基）的连接增强了翻译，如通过体外和体内翻译测定所评估的。YB-1 5 '-UTR的缺失构建体也导致更高的翻译效率，特别是在从主要转录起始位点定位到+197至+331的区域中。RNA凝胶迁移实验表明，YB-1对各种5 '-UTR探针序列的亲和力对于全长5'-UTR比对于缺失的5 '-UTR序列更高。体外翻译实验证明重组YB-1蛋白可抑制YB-1 mRNA全长5 '-UTR的翻译。因此，我们的研究结果为YB-1 mRNA翻译通过5 '-UTR的自动调节提供了证据。
The eukaryotic Y-box binding protein YB-1 is involved in various biological processes, including DNA repair, cell proliferation and the regulation of transcription and translation. YB-1 protein is abundant and expressed ubiquitously in human cells, functioning in cell proliferation and transformation. Its concentration is thought to be highly regulated at both the levels of transcription and translation. Therefore, we investigated whether or not the 5'-UTR of YB-1 mRNA affects the translation of YB-1 protein, thus influencing expression levels. Luciferase mRNA ligated to the YB-1 mRNA 5'-UTR was used as a reporter construct. Ligation of the full-length YB-1 5'-UTR (331 bases) enhanced translation as assessed by in vitro and in vivo translation assays. Deletion constructs of the YB-1 5'-UTR also resulted in a higher efficiency of translation, especially in the region mapped to +197 to +331 from the major transcription start site. RNA gel shift assays revealed that the affinity of YB-1 for various 5'-UTR probe sequences was higher for the full-length 5'-UTR than for deleted 5'-UTR sequences. An in vitro translation assay was used to demonstrate that recombinant YB-1 protein inhibited translation of the full-length 5'-UTR of YB-1 mRNA. Thus, our findings provide evidence for the autoregulation of YB-1 mRNA translation via the 5'-UTR.