Natural trans-spliced mRNAs are generated from the human estrogen receptor-α (hERα) gene
Natural trans-spliced mRNAs are generated from the human estrogen receptor-α (hERα) gene
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DOI：
10.1074/jbc.m203513200
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发表时间：
2002-07-19
影响因子：
4.8
通讯作者：
Gannon, F
中科院分区：
文献类型：
--
作者：
Flouriot, G;Brand, H;Gannon, F
人类雌激素受体- α (herα)基因是一个复杂的基因组单位，以组织特异性的方式表现出选择性剪接和启动子的使用。在通过快速扩增5‘ cDNA末端来研究新的hERalpha mRNA变异体的过程中，我们发现了一个cDNA，其中外显子1A的受体位点（不同的领导外显子通常被交替剪接到其中）准确地剪接在外显子1A的3’末端（打乱1A- >1A hERalpha cDNA）。逆转录pcr和S1核酸酶定位分析显示，1A—>1A hERalpha转录本不是仅由1A外显子组成的环状rna，而是对应于由8个hERalpha基因编码外显子组成的线性多腺苷化的hERalpha rna，其特征是1A外显子的重复。基因组Southern blot实验排除了人类基因组中hERalpha外显子1A重复的假设。因此，这些数据表明1A—>1A hERalpha转录本可能是通过反式剪接产生的。在瞬时转染实验中证明，通过反式剪接由一个hERalpha外显子1A、外显子2及其侧面内含子区域组成的嵌合基因产生的pre- mrna产生了这种转录本。因此，除了选择性的顺式剪接，herα基因也受到自然反式剪接的影响。
The human estrogen receptor-alpha (heralpha) gene is a complex genomic unit exhibiting alternative splicing and promoter usage in a tissue-specific manner. During the investigation of new hERalpha mRNA variants by rapid amplification of 5' cDNA ends, we identified a cDNA in which the acceptor site of exon 1A, into which the different leader exons are normally alternatively spliced, was spliced accurately the 3' extremity of exon 1A (scrambled 1A-->1A hERalpha cDNA). Reverse transcription-PCR and S1 nuclease mapping analysis revealed that 1A-->1A hERalpha transcripts were not circular RNAs constituted by exon 1A only but corresponded to linear polyadenylated hERalpha RNAs composed of the eight coding exons of the hERalpha gene and characterized by a duplication of exon 1A. Genomic Southern blot experiments excluded the hypothesis of duplication of hERalpha exon 1A in the human genome. Therefore, these data suggested that 1A-->1A hERalpha transcripts were likely generated by trans-splicing. The production of such transcripts by trans-splicing of pre-mRNAs generated from a chimeric gene formed by a single hERalpha exon 1A, exon 2, and their flanking intronic regions was demonstrated in transient transfection experiments. Therefore, in addition to the alternative cis-splicing, the hERalpha gene is also subject to natural trans-splicing.