Binding specificity and mRNA targets of a C-elegans PUF protein, FBF-1
Binding specificity and mRNA targets of a C-elegans PUF protein, FBF-1
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DOI：
10.1261/rna.7255805
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发表时间：
2005-04-01
期刊：
影响因子：
4.5
通讯作者：
Wickens, M
中科院分区：
文献类型：
--
作者：
Bernstein, D;Hook, B;Wickens, M
序列特异性RNA-蛋白质相互作用是许多mRNA调控的基础。在这里，我们分析RNA序列特异性秀丽隐杆线虫FBF-1，PUF蛋白家族的创始成员。与其他PUF蛋白一样，FRF-1与靶mRNA的3' UTR结合并降低这些靶基因的表达。在这里，我们表明，FBF-1和它的近亲，FBF-2，结合具有相似的亲和力，多个RNA位点。我们使用诱变和体内选择实验来鉴定对FBF-1结合至关重要的核苷酸。结合元件包含“核心”中心区和侧翼序列。核心区域与其他PUF蛋白的结合位点相似但不同。我们联合收割机将结合元件的鉴定与信息学相结合来预测一个C. elegans 3' UTR数据库。这些数据鉴定了FBF-1和FBF-2的一组新的候选mRNA靶标。
Sequence-specific RNA-protein interactions underlie regulation of many mRNAs. Here we analyze the RNA sequence specificity of Caenorhabditis elegans FBF-1, a founding member of the PUF protein family. Like other PUF proteins, FRF-1 binds to the 3' UTR of target mRNAs and decreases expression of those target genes. Here, we show that FBF-1 and its close relative, FBF-2, bind with similar affinity to multiple RNA sites. We use mutagenesis and in vivo selection experiments to identify nucleotides that are essential for FBF-1 binding. The binding elements comprise a "core" central region and flanking sequences. The core region is similar but distinct from the binding sites of other PUF proteins. We combine the identification of binding elements with informatics to predict new FBF-1 binding sites in a C. elegans 3' UTR database. These data identify a set of new candidate mRNA targets of FBF-1 and FBF-2.