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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Genome-wide identification of mRNAs associated with the translational regulator PUMILIO in Drosophila melanogaster
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Genome-wide identification of mRNAs associated with the translational regulator PUMILIO in Drosophila melanogaster

机译：全基因组识别与果蝇果蝇中翻译调节因子PUMILIO相关的mRNA

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摘要

Genome-wide identification of RNAs associated with RNA-binding proteins is crucial for deciphering posttranscriptional regulatory systems. PUMILIO is a member of the evolutionary conserved Puf-family of RNA-binding proteins that repress gene expression posttranscriptionally. We generated transgenic flies expressing affinity-tagged PUMILIO under the control of an ovary-specific promoter, and we purified PUMILIO from whole adult flies and embryos and analyzed associated mRNAs by using DNA microar-rays. Distinct sets comprising hundreds of mRNAs were associated with PUMILIO at the two developmental stages. Many of these mRNAs encode functionally related proteins, supporting a model for coordinated regulation of posttranscriptional modules by specific RNA-binding proteins. We identified a characteristic sequence motif in the 3′-untranslated regions of mRNAs associated with PUMILIO, and the sufficiency of this motif for interaction with PUMILIO was confirmed by RNA pull-down experiments with biotinylated synthetic RNAs. The RNA motif strikingly resembles the one previously identified for Puf3p, one of five Saccharomyces cerevisiae Puf proteins; however, proteins encoded by the associated mRNAs in yeast and Drosophila do not appear to be related. The results suggest extensive posttranscriptional regulation by PUMILIO and uncover evolutionary features of this conserved family of RNA-binding proteins.
机译：与RNA结合蛋白相关的RNA的全基因组鉴定对于解密转录后调控系统至关重要。 PUMILIO是RNA结合蛋白进化保守的Puf家族的成员，该家族在转录后抑制基因表达。我们在卵巢特异性启动子的控制下产生了表达亲和标记的PUMILIO的转基因果蝇，并从整个成年果蝇和胚胎中纯化了PUMILIO，并使用DNA微阵列分析了相关的mRNA。在两个发育阶段，包含数百种mRNA的不同集合与PUMILIO相关。这些mRNA中的许多都编码功能相关的蛋白质，从而支持通过特异性RNA结合蛋白对转录后模块进行协调调控的模型。我们在与PUMILIO相关的mRNA的3'-非翻译区中鉴定了一个特征序列基序，并且该基序与PUMILIO相互作用的充分性已通过生物素化合成RNA的RNA下拉实验得到证实。 RNA的基序非常类似于先前针对Puf3p鉴定的5个酿酒酵母Puf蛋白之一。然而，酵母和果蝇中由相关的mRNA编码的蛋白质似乎并不相关。结果表明由PUMILIO进行广泛的转录后调控，并揭示了这种保守的RNA结合蛋白家族的进化特征。

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