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一种植物U1 snRNP 70K基因的结构与表达:U1 snRNP 70K前体mRNA的可变剪接产生两种不同的转录本。

Structure and expression of a plant U1 snRNP 70K gene: alternative splicing of U1 snRNP 70K pre-mRNAs produces two different transcripts.

作者信息

Golovkin M, Reddy A S

机构信息

Department of Biology, Colorado State University, Fort Collins 80523, USA.

出版信息

Plant Cell. 1996 Aug;8(8):1421-35. doi: 10.1105/tpc.8.8.1421.

Abstract

The product of the U1 small nuclear ribonucleoprotein particle (U1 snRNP) 70K (U1-70K) gene, a U1 snRNP-specific protein, has been implicated in basic as well as alternative splicing of pre-mRNAs in animals. Here, we report the isolation of full-length cDNAs and the corresponding genomic clone encoding a U1-70K protein from a plant system. The Arabidopsis U1-70K protein is encoded by a single gene, which is located on chromosome 3. Several lines of evidence indicate that two distinct transcripts (short and long) are produced from the same gene by alternative splicing of the U1-70K pre-mRNA. The alternative splicing involves inclusion or exclusion of a region (910 bp) that we named "included intron." Two transcripts were clearly detectable in all tissues tested, and the level of the transcripts varied in different organs. The deduced amino acid (427 residues) sequence from the short transcript has strong homology to the animal U1-70K protein and contains an RNA recognition motif, a glycine hinge, and an arginine-rich region characteristic of the animal U1-70K protein. The long transcript has an in-frame translational termination codon within the 910-bp included intron, resulting in a truncated protein containing only 204 amino acids. The protein encoded by the short transcript is recognized by U1 RNP-specific monoclonal antibodies and binds specifically to the Arabidopsis U1 snRNA, whereas the protein from the long transcript does not. In addition, multiple polyadenylation sites were observed in the 3' untranslated region. These results suggest a complex post-transcriptional regulation of Arabidopsis U1-70K gene expression.

摘要

U1小核核糖核蛋白颗粒(U1 snRNP)70K(U1-70K)基因的产物是一种U1 snRNP特异性蛋白,已被证明与动物前体mRNA的基础剪接和可变剪接有关。在此,我们报告了从植物系统中分离出全长cDNA以及编码U1-70K蛋白的相应基因组克隆。拟南芥U1-70K蛋白由单个基因编码,该基因位于3号染色体上。几条证据表明,通过U1-70K前体mRNA的可变剪接,同一基因产生了两种不同的转录本(短转录本和长转录本)。可变剪接涉及一个我们命名为“包含内含子”的区域(910 bp)的包含或排除。在所有测试组织中都能清楚地检测到两种转录本,并且转录本的水平在不同器官中有所不同。从短转录本推导的氨基酸序列(427个残基)与动物U1-70K蛋白具有高度同源性,并且包含一个RNA识别基序、一个甘氨酸铰链和一个动物U1-70K蛋白特有的富含精氨酸的区域。长转录本在910 bp的包含内含子内有一个框内翻译终止密码子,导致产生仅包含204个氨基酸的截短蛋白。短转录本编码的蛋白被U1 RNP特异性单克隆抗体识别,并特异性结合拟南芥U1 snRNA,而长转录本的蛋白则不能。此外,在3'非翻译区观察到多个聚腺苷酸化位点。这些结果表明拟南芥U1-70K基因表达存在复杂的转录后调控。

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