Ribonomic analysis of human Pum1 reveals cis-trans conservation across species despite evolution of diverse mRNA target sets
Ribonomic analysis of human Pum1 reveals cis-trans conservation across species despite evolution of diverse mRNA target sets
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DOI：
10.1128/mcb.00155-08
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发表时间：
2008-06-01
影响因子：
5.3
通讯作者：
Keene, Jack D.
中科院分区：
文献类型：
--
作者：
Morris, Adam R.;Mukherjee, Neelanjan;Keene, Jack D.
PUF 家族蛋白是非哺乳动物物种中特征最清楚的调节性 RNA 结合蛋白之一，但人们对哺乳动物 PUF 蛋白的 mRNA 靶标或功能知之甚少。在这项研究中，我们使用核糖组分析来识别和分析与含有内源性人类 PUF 蛋白 Pum1 的核糖核蛋白相关的 mRNA。 Pum1 相关 mRNA 高度富集编码在转录调节和细胞周期/增殖中发挥作用的蛋白质的基因，结果与转录后 RNA 调节子模型以及 PUF 蛋白质在干细胞生物学中的祖先功能一致。对 Pum1 相关 mRNA 的 3' 非翻译区序列的分析揭示了核心 Pum1 共有序列 UGUAHAUA。 Pum1 敲低表明 Pum1 增强了相关 mRNA 的衰变，Pum1 重新定位到应激颗粒表明 Pum1 在抑制翻译中发挥作用。这项研究是哺乳动物 PUF 蛋白的首次体内全基因组 mRNA 靶标鉴定，并提供了人类 PUF 蛋白调节相关 mRNA 稳定性的直接证据。将 Pum1 相关 mRNA 与来自酿酒酵母和黑腹果蝇的 PUF 蛋白 mRNA 靶标进行比较，证明了保守的 RNA 结合结构域和同源结合序列如何在进化上重新连接，以调节不同组功能相关基因的集体表达。
PUF family proteins are among the best-characterized regulatory RNA-binding proteins in nonmammalian species, but relatively little is known about mRNA targets or functions of mammalian PUF proteins. In this study, we used ribonomic analysis to identify and analyze mRNAs associated with ribonucleoproteins containing an endogenous human PUF protein, Pum1. Pum1-associated mRNAs were highly enriched for genes encoding proteins that function in transcriptional regulation and cell cycle/proliferation, results consistent with the posttranscriptional RNA regulon model and the proposed ancestral functions of PUF proteins in stem cell biology. Analysis of 3' untranslated region sequences of Pum1-associated mRNAs revealed a core Pum1 consensus sequence, UGUAHAUA. Pum1 knockdown demonstrated that Pum1 enhances decay of associated mRNAs, and relocalization of Pum1 to stress granules suggested that Pum1 functions in repression of translation. This study is the first in vivo genome-wide mRNA target identification of a mammalian PUF protein and provides direct evidence that human PUF proteins regulate stability of associated mRNAs. Comparison of Pum1-associated mRNAs to mRNA targets of PUF proteins from Saccharomyces cerevisiae and Drosophila melanogaster demonstrates how a well-conserved RNA-binding domain and cognate binding sequence have been evolutionarily rewired to regulate the collective expression of different sets of functionally related genes.