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Target specificity of neuronal RNA-binding protein, Mel-N1: direct binding to the 3' untranslated region of its own mRNA.

机译:神经元RNA结合蛋白Mel-N1的靶标特异性:直接结合至其自身mRNA的3'非翻译区。

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摘要

We have identified cDNAs encoding Mel-N1, the mouse homologue of a human nervous system-specific RNA-binding protein, Hel-N1. Two major mRNA transcripts of Mel-N1 were detected predominantly in the adult mouse brain by Northern blot analysis. To gain insight into the RNA binding specificity of Mel-N1, we performed iterative in vitro RNA selection. The resulting in vitro selected RNAs were found to contain AU-rich sequences as well as a GAAA motif in the majority of clones. By means of in vitro binding assays we demonstrate that this GAAA sequence appears to significantly affect the Mel-N1 RNA-binding efficiency. Our studies further reveal that Mel-N1 can bind to its own 3' untranslated region (3'UTR) as well as to the c-fos 3'UTR, and is localized predominantly in the cytoplasmic region in cells, suggesting that posttranscriptional autoregulation of Mel-N1 gene expression occurs in vivo.
机译:我们已经确定了编码Mel-N1的cDNA,Mel-N1是人类神经系统特异性RNA结合蛋白Hel-N1的小鼠同源物。通过RNA印迹分析,主要在成年小鼠脑中检测到Mel-N1的两个主要mRNA转录物。为了深入了解Mel-N1的RNA结合特异性,我们进行了体外迭代RNA选择。发现所得的体外选择的RNA在大多数克隆中均含有富含AU的序列以及GAAA基序。通过体外结合测定,我们证明了该GAAA序列似乎显着影响Mel-N1 RNA结合效率。我们的研究进一步揭示,Mel-N1可以与其自身的3'非翻译区(3'UTR)以及c-fos 3'UTR结合,并且主要位于细胞的细胞质区域,这表明该蛋白的转录后自动调节Mel-N1基因表达发生在体内。

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