cDNA structure, expression and nucleic acid-binding properties of three RNA-binding proteins in tobacco: occurrence of tissue-specific alternative splicing.
cDNA structure, expression and nucleic acid-binding properties of three RNA-binding proteins in tobacco: occurrence of tissue-specific alternative splicing.
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烟草中三种RNA结合蛋白的cDNA结构、表达和核酸结合特性：组织特异性选择性剪接的发生。
DOI：
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发表时间：
1993
影响因子：
14.9
通讯作者：
Masahiro Sugiura
中科院分区：
文献类型：
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作者：
T. Hirose;Mamoru Sugita;Masahiro Sugiura
从烟草（Nicotiana sylvestris）cDNA文库中分离出三个编码RNA结合蛋白的cDNA。预测的蛋白质（RGP-1）彼此同源，并且由N-末端一半中的80个氨基酸的共有序列型RNA结合结构域和C-末端一半中的61-78个氨基酸的富含甘氨酸的结构域组成。利用体外合成的RGP-1蛋白进行核酸结合试验，证实它是一种RNA结合蛋白。基于其对聚（G）和聚（U）的强亲和力，RGP-1蛋白被认为特异性结合富含G和/或U的序列。所有这三个基因在叶、根、花和培养细胞中表达，然而，大量的前体mRNA积累，特别是在根中。序列分析和核糖核酸酶保护试验表明，大量的选择性剪接的mRNA，这是由5'剪接位点的差异选择，也存在于各种组织。在这三个基因中的两个中发现了组织特异性选择性剪接。选择性剪接的mRNA也在多核糖体组分中检测到，并被认为产生截短的多肽。这种选择性剪接的可能作用进行了讨论。
Three cDNAs encoding RNA-binding proteins were isolated from a tobacco (Nicotiana sylvestris) cDNA library. The predicted proteins (RGP-1) are homologous to each other and consist of a consensus-sequence type RNA-binding domain of 80 amino acids in the N-terminal half and a glycine-rich domain of 61-78 amino acids in the C-terminal half. Nucleic acid-binding assay using the in vitro synthesized RGP-1 protein confirmed that it is an RNA-binding protein. Based on its strong affinity for poly(G) and poly(U), the RGP-1 proteins are suggested to bind specifically to G and/or U rich sequences. All three genes are expressed in leaves, roots, flowers and cultured cells, however, the substantial amount of pre-mRNAs are accumulated especially in roots. Sequence analysis and ribonuclease protection assay indicated that significant amounts of alternatively spliced mRNAs, which are produced by differential selection of 5' splice sites, are also present in various tissues. Tissue-specific alternative splicing was found in two of the three genes. The alternatively spliced mRNAs are also detected in polysomal fractions and are suggested to produce truncated polypeptides. A possible role of this alternative splicing is discussed.