热休克刺激通过转录后机制调控NFKBIA基因的表达
Heat shock treatment regulates the expression of NFKBIA (IκBα) in a post-transcriptional manner
由NFKBIA基因编码表达的IκB蛋白是最早被鉴定的转录因子NF-κB的抑制蛋白，而其转录受NF-κB的直接调控的机制已经研究得很清楚，但是转录后机制调控其在细胞中表达的机制目前仍不清楚。为了揭示NFKBIA基因表达的转录后调控机制，我们首先将源自不同种属（包括人、小鼠、大鼠、狗与牛）的NFKBIA mRNA的3?-UTR采用Clustal Omega多序列对位排列软进行分析，结果显示其在这些被分析的种属之中高度保守。进一步采用双荧光素酶报告基因表达系统分析发现小鼠NFKBIA 3?-UTR可下调与其相连的荧光素报告基因的表达，而热休克处理可以促进与其相连的荧光素酶报告基因的表达，并且可提高NFKBIA mRNA的稳定性。采用生物标记RNA pulldown实验结合质谱分析技术鉴定了与 小鼠NFKBIA mRNA 3?-UTR特异性结合的蛋白质，其中一半为组成核糖体40S与60S亚基的核糖体蛋白。
IκBα, encoded by NFKBIA, is the prototype inhibitor of transcription factor NF-κB. Notably, the transcription of NFKBIA is directly regulated by NF-κB. In contrast, the contribution of post-transcriptional regulation in the expression of NFKBIA remains obscure. To elucidate the post-transcriptional regulation of NFKBIA, we first aligned the 3?-UTR from several different species (human, mouse, rat, dog and cow). The alignment results revealed that the 3?-UTR sequences were highly conserved among these analyzed species. Dual luciferase assay analysis showed that 3?-UTR of mouse NFKBIA downregulated the expression of associated luciferase gene. Heat shock treatment stabilized NFKBIA mRNA and promote the expression of luciferase gene associated with mouse NFKBIA 3?-UTR. Biotin-labeled RNA pulldown assay followed mass spectrometry analysis identified specific binding proteins to the 3?-UTR of mouse NFKBIA mRNA. Half of them were identified as ribosomal proteins of 40S and 60S ribosome subunits.
欧小利、陈新玉、姜勇、梅柱中、王妮、李涛
分子生物学细胞生物学生物化学
NFKBIA转录后调控RNA结合蛋白
NFKBIApost-transcriptional regulationRNA binding protein
欧小利,陈新玉,姜勇,梅柱中,王妮,李涛.热休克刺激通过转录后机制调控NFKBIA基因的表达[EB/OL].(2014-01-15)[2025-07-02].http://www.paper.edu.cn/releasepaper/content/201401-642.点此复制
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