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首页> 外文期刊>Nucleic Acids Research >A novel cis-acting element from the 3'UTR of DNA damage-binding protein 2 mRNA links transcriptional and post-transcriptional regulation of gene expression
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A novel cis-acting element from the 3'UTR of DNA damage-binding protein 2 mRNA links transcriptional and post-transcriptional regulation of gene expression

机译：DNA损伤结合蛋白2 mRNA 3'UTR产生的新型顺式作用元件连接基因表达的转录和转录后调控

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摘要

The DNA damage-binding protein 2 (DDB2) is an adapter protein that can direct a modular Cul4-DDB1-RING E3 Ligase complex to sites of ultraviolet light-induced DNA damage to ubiquitinate substrates during nucleotide excision repair. The DDB2 transcript is ultraviolet-inducible; therefore, its regulation is likely important for its function. Curiously, the DDB2 mRNA is reportedly short-lived, but the transcript does not contain any previously characterized cis-acting determinants of mRNA stability in its 3' untranslated region (3'UTR). Here, we used a tetracycline regulated d2EGFP reporter construct containing specific 3'UTR sequences from DDB2 to identify novel cis-acting elements that regulate mRNA stability. Synthetic 3'UTRs corresponding to sequences as short as 25 nucleotides from the central region of the 3'UTR of DDB2 were sufficient to accelerate decay of the heterologous reporter mRNA. Conversely, these same 3'UTRs led to more rapid induction of the reporter mRNA, export of the message to the cytoplasm and the subsequent accumulation of the encoded reporter protein, indicating that this newly identified cis-acting element affects transcriptional and post-transciptional processes. These results provide clear evidence that nuclear and cytoplasmic processing of the DDB2 mRNA is inextricably linked.
机译：DNA损伤结合蛋白2（DDB2）是一种衔接蛋白，可在核苷酸切除修复过程中将模块化Cul4-DDB1-RING E3连接酶复合物引导至紫外线诱导的DNA损伤泛素化底物的位点。 DDB2转录本是紫外线诱导的;因此，其调节对其功能可能很重要。奇怪的是，据报道DDB2 mRNA寿命短，但该转录本在其3'非翻译区（3'UTR）中不包含任何先前表征的mRNA稳定性的顺式决定簇。在这里，我们使用了四环素调节的d2EGFP报告基因构建体，该构建体包含来自DDB2的特定3'UTR序列，以鉴定可调节mRNA稳定性的新型顺式作用元件。对应于距DDB2 3'UTR中心区域短至25个核苷酸的序列的合成3'UTR足以加速异源报道基因mRNA的降解。相反，这些相同的3'UTR导致报告基因mRNA的更快诱导，信息向细胞质的输出以及随后编码的报告蛋白的积累，表明这种新近鉴定的顺式作用元件影响转录和转录后过程。 。这些结果提供了清楚的证据，表明DDB2 mRNA的核和细胞质加工紧密相关。

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