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首页> 美国卫生研究院文献>The Journal of Biological Chemistry >The Drosophila Tis11 Protein and Its Effects on mRNA Expression in Flies
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The Drosophila Tis11 Protein and Its Effects on mRNA Expression in Flies

机译：果蝇Tis11蛋白及其对果蝇mRNA表达的影响。

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摘要

Members of the mammalian tristetraprolin family of CCCH tandem zinc finger proteins can bind to certain AU-rich elements (AREs) in mRNAs, leading to their deadenylation and destabilization. Mammals express three or four members of this family, but Drosophila melanogaster and other insects appear to contain a single gene, Tis11. We found that recombinant Drosophila Tis11 protein could bind to ARE-containing RNA oligonucleotides with low nanomolar affinity. Remarkably, co-expression in mammalian cells with “target” RNAs demonstrated that Tis11 could promote destabilization of ARE-containing mRNAs and that this was partially dependent on a conserved C-terminal sequence resembling the mammalian NOT1 binding domain. Drosophila Tis11 promoted both deadenylation and decay of a target transcript in this heterologous cell system. We used chromosome deletion/duplication and P element insertion to produce two types of Tis11 deficiency in adult flies, both of which were viable and fertile. To address the hypothesis that Tis11 deficiency would lead to the abnormal accumulation of potential target transcripts, we analyzed gene expression in adult flies by deep mRNA sequencing. We identified 69 transcripts from 56 genes that were significantly up-regulated more than 1.5-fold in both types of Tis11-deficient flies. Ten of the up-regulated transcripts encoded probable proteases, but many other functional classes of proteins were represented. Many of the up-regulated transcripts contained potential binding sites for tristetraprolin family member proteins that were conserved in other Drosophila species. Tis11 is thus an ARE-binding, mRNA-destabilizing protein that may play a role in post-transcriptional gene expression in Drosophila and other insects.
机译：CCCH串联锌指蛋白的哺乳动物tristetraprolin家族成员可以与mRNA中的某些富含AU的元件（ARE）结合，从而导致它们的腺苷酸化和去稳定作用。哺乳动物表达了这个家族的三到四个成员，但是果蝇和其他昆虫似乎只包含一个基因Tis11。我们发现，果蝇Tis11重组蛋白可以低纳摩尔亲和力与含ARE的RNA寡核苷酸结合。值得注意的是，在哺乳动物细胞中与“靶标” RNA的共表达表明Tis11可以促进含ARE的mRNA的不稳定，并且这部分依赖于类似于哺乳动物NOT1结合域的保守C端序列。果蝇Tis11在该异源细胞系统中促进了腺苷酸化和目标转录物的降解。我们使用染色体缺失/复制和P元素插入来在成年果蝇中产生两种类型的Tis11缺乏症，两者都是可行的和可育的。为了解决Tis11缺乏会导致潜在目标转录本异常积累的假设，我们通过深mRNA测序分析了成年果蝇的基因表达。我们从56种基因中鉴定出69个转录本，在两种Tis11缺陷型果蝇中均显着上调了1.5倍以上。十个上调的转录本编码可能的蛋白酶，但代表了许多其他功能的蛋白质。许多上调的转录本包含潜在的结合Tristetraprolin家族成员蛋白的位点，在其他果蝇物种中是保守的。因此，Tis11是一种ARE结合，mRNA不稳定的蛋白，可能在果蝇和其他昆虫的转录后基因表达中发挥作用。

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