The calcitonin exon and its flanking intronic sequences are sufficient for the regulation of human calcitonin/calcitonin gene-related peptide alternative RNA splicing.
The calcitonin exon and its flanking intronic sequences are sufficient for the regulation of human calcitonin/calcitonin gene-related peptide alternative RNA splicing.
复制标题
DOI：
10.1210/mend.8.12.7535892
复制
发表时间：
1994-12
影响因子：
--
通讯作者：
H. Lou;G. Cote;R. Gagel
中科院分区：
文献类型：
--
作者：
H. Lou;G. Cote;R. Gagel
降钙素（CT）/降钙素基因相关肽（CGRP）的初级转录物以细胞特异性方式选择性剪接，以在甲状腺C细胞中产生CT，在神经元细胞中产生CGRP。这一调控过程的关键步骤是识别和包含外显子4产生CT mRNA或不识别和排除外显子4产生CGRP mRNA。为了确定CT外显子的包含/排除是否独立于其位置而受到调节，我们创建了一系列含有递减量的CT基因序列的小基因构建体。测试了人胶质母细胞瘤细胞系T98 G并将其用作CT外显子排除细胞系，而HeLa细胞用作CT外显子包含细胞系。当CT基因内外显子4的相对位置改变或当具有侧翼序列的外显子插入到完全异源的基因中时，CT外显子包含/排除受到调节。我们的研究结果表明，CT外显子功能作为一个单位，在一个位置独立的方式调节自己的包容/排斥。我们相信，异源融合基因只包含外显子4和其侧翼内含子序列的一部分，将有助于进一步确定参与调节CT/CGRP剪接的序列元件。
The primary transcript of the calcitonin (CT)/calcitonin gene-related peptide (CGRP) is alternatively spliced in a cell-specific fashion to produce CT in thyroid C cells and CGRP in neuronal cells. The key step in this regulatory process is the recognition and inclusion of exon 4 to produce CT mRNA or nonrecognition and exclusion of exon 4 to produce CGRP mRNA. To determine whether inclusion/exclusion of CT exon is regulated independently of its position, we created a series of minigene constructs containing decreasing amounts of CT gene sequence. A human glioblastoma cell line, T98G, was tested and used as a CT exon exclusion cell line, while HeLa cells were used as a CT exon inclusion cell line. CT exon inclusion/exclusion was regulated when either the relative position of exon 4 within the CT gene was changed or when the exon with flanking sequence was inserted into a completely heterologous gene. Our results demonstrate that CT exon functions as a unit in a position-independent fashion in regulating its own inclusion/exclusion. We believe that the heterologous fusion gene containing only exon 4 and part of its flanking intron sequences will be useful for further defining the sequence elements involved in the regulation of CT/CGRP splicing.