A non-sequence-specific double-stranded RNA structural element regulates splicing of two mutually exclusive exons of fibroblast growth factor receptor 2 (FGFR2)
A non-sequence-specific double-stranded RNA structural element regulates splicing of two mutually exclusive exons of fibroblast growth factor receptor 2 (FGFR2)
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DOI：
10.1074/jbc.m207409200
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发表时间：
2002-12-20
影响因子：
4.8
通讯作者：
Carstens, RP
中科院分区：
文献类型：
--
作者：
Muh, SJ;Hovhannisyan, RH;Carstens, RP
成纤维细胞生长因子受体2 （FGFR2）互斥外显子IIIb和IIIc的选择性剪接代表了转录后基因调控的一个严格调控和功能相关的例子。大鼠前列腺癌DT3和AT3细胞系分别显示出IIIb或IIIc外显子的排他性选择，并已被用于表征剪接调节所需的FGFR2 RNA顺式元件。两个被称为ISE-2和ISAR的序列位于IIIb和IIIc外显子之间的内含子中，是细胞类型特异性外显子IIIb所必需的。先前的研究表明，这些元素的功能涉及RNA茎结构的形成，尽管它们之间相隔700多个核苷酸。使用转染的minigenes，我们对ISE-2和ISAR的序列和结构成分进行了系统的分析，这些序列和结构成分是调节FGFR2剪接能力所必需的。我们发现这些元件的一级序列可以被完全不相关的序列所取代，只要它们也被预测形成RNA茎结构。因此，非序列特异性双链RNA干构成了FGFR2剪接所需的功能元件，这表明双链RNA结合蛋白是剪接调节机制的一个组成部分。
Alternative splicing of fibroblast growth factor receptor 2 (FGFR2) mutually exclusive exons IIIb and IIIc represents a tightly regulated and functionally relevant example of post-transcriptional gene regulation. Rat prostate cancer DT3 and AT3 cell lines demonstrate exclusive selection of either exon IIIb or exon IIIc, respectively, and have been used to characterize regulatory FGFR2 RNA cis-elements that are required for splicing regulation. Two sequences termed ISE-2 and ISAR are located in the intron between exons IIIb and IIIc and are required for cell-type specific exon IIIb. Previous studies suggest that the function of these elements involves formation of an RNA stem structure, even though they are separated by more than 700 nucleotides. Using transfected minigenes, we performed a systematic analysis of the sequence and structural components of ISE-2 and ISAR that are required for their ability to regulate FGFR2 splicing. We found that the primary sequence of these elements can be replaced by completely unrelated sequences, provided that they are also predicted to form an RNA stem structure. Thus, a nonsequence-specific double stranded RNA stem constitutes a functional element required for FGFR2 splicing, suggesting that a double-stranded RNA binding protein is a component of the splicing regulatory machinery.