Novel RNA-binding activity of NQ01 promotes SERPINA1 mRNA translation
Novel RNA-binding activity of NQ01 promotes SERPINA1 mRNA translation
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DOI：
10.1016/j.freeradbiomed.2016.08.005
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发表时间：
2016-10-01
影响因子：
7.4
通讯作者：
de Cabo, Rafael
中科院分区：
文献类型：
--
作者：
Di Francesco, Andrea;Di Germanio, Clara;de Cabo, Rafael
NAD(P)H：醌氧化还原酶 (NQO1) 对于细胞防御活性氧化物质、癌症和代谢应激至关重要。最近，在核糖核蛋白 (RNP) 复合物中发现了 NQO1，但与 NQO1 相互作用的 mRNA 以及此类相互作用的功能影响尚不清楚。在这里，我们使用核糖核蛋白免疫沉淀（RIP）和微阵列分析来全面鉴定人肝癌 HepG2 细胞中的 NQOI 靶 mRNA 子集。其主要靶标之一 SERPINAI mRNA 编码丝氨酸蛋白酶抑制剂 oc-1-抗胰蛋白酶 (AlAT)，它与肥胖相关代谢炎症、慢性阻塞性肺病 (COPD)、肝硬化和肝细胞癌等疾病相关。生物素下拉分析表明NQO1可以结合SERPINAI mRNA的3'非翻译区(UTR)和编码区(CR)。 NQOI 不影响 SERPINAI mRNA 水平；相反，它增强了 SERPINAI mRNA 的翻译，因为 NQOI 沉默减少了 SERPINAI mRNA 上形成的多核糖体的大小，并降低了 AlAT 的丰度。荧光素酶报告基因分析进一步表明，NQOI 通过 SERPINAI 3'UTR 调节 SERPINAI mRNA 翻译。因此，NQO1-KO 小鼠的肝脏和血清 AlAT 水平降低，中性粒细胞弹性蛋白酶 (NE) 活性增加，NE 是 AlAT 的主要靶标之一。我们认为 NQOI 作为 RNA 结合蛋白的这种新作用机制可能有助于解释其多效性生物学效应。由爱思唯尔公司出版
NAD(P)H: quinone oxidoreductase (NQO1) is essential for cell defense against reactive oxidative species, cancer, and metabolic stress. Recently, NQO1 was found in ribonucleoprotein (RNP) complexes, but NQO1-interacting mRNAs and the functional impact of such interactions are not known. Here, we used ribonucleoprotein immunoprecipitation (RIP) and microarray analysis to identify comprehensively the subset of NQOI target mRNAs in human hepatoma HepG2 cells. One of its main targets, SERPINAI mRNA, encodes the serine protease inhibitor oc-1-antitrypsin, AlAT, which is associated with disorders including obesity-related metabolic inflammation, chronic obstructive pulmonary disease (COPD), liver cirrhosis and hepatocellular carcinoma. Biotin pulldown analysis indicated that NQO1 can bind the 3' untranslated region (UTR) and the coding region (CR) of SERPINAI mRNA. NQOI did not affect SERPINAI mRNA levels; instead, it enhanced the translation of SERPINAI mRNA, as NQOI silencing decreased the size of polysomes forming on SERPINAI mRNA and lowered the abundance of AlAT. Luciferase reporter analysis further indicated that NQOI regulates SERPINAI mRNA translation through the SERPINAI 3'UTR. Accordingly, NQO1-KO mice had reduced hepatic and serum levels of AlAT and increased activity of neutrophil elastase (NE), one of the main targets of AlAT. We propose that this novel mechanism of action of NQOI as an RNA-binding protein may help to explain its pleiotropic biological effects. Published by Elsevier Inc.