Alternative promoter usage and alternative splicing of the rat estrogen receptor α gene generate numerous mRNA variants with distinct 5′-ends
Alternative promoter usage and alternative splicing of the rat estrogen receptor α gene generate numerous mRNA variants with distinct 5′-ends
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DOI：
10.1016/j.jsbmb.2009.10.001
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发表时间：
2010-01-01
影响因子：
4.1
通讯作者：
Sakuma, Yasuo
中科院分区：
文献类型：
--
作者：
Ishii, Hirotaka;Kobayashi, Momoko;Sakuma, Yasuo
雌激素受体α（ER α）基因的T非翻译区（UTR）在决定组织特异性表达中起重要作用。为了阐明大鼠ER α基因表达的调控机制，必须对基因组结构进行研究。因此，我们使用5 '-cDNA末端快速扩增（5'-RACE）和RT-PCR分析了大鼠ER α mRNA 5 '-UTR的结构。分析显示存在含有独特5 '-UTR的多种变体。我们绘制了大鼠基因组上的cDNA序列，并新鉴定了一个前导外显子（外显子0 U）和十个非翻译的内部外显子（外显子11-10）。从四个不同的前导外显子（外显子0 S、0 N、0 U和0/B）剪接到外显子1和与十一个内部外显子（外显子I1-10和OT）组合的选择性剪接产生多个转录物。RT-PCR分析表明，每个变体有优选的表达位点，这表明启动子的使用和剪接的组织特异性的方式进行调节。此外，我们确定了产生Delta外显子1变体（0 S-2-3-4-5-6-7-8）的剪接事件，其被翻译成大鼠46 kDa ER α蛋白。我们的研究结果表明，大鼠ER α基因是更复杂的比以前认为的基因组组织和选择性启动子的使用和选择性剪接的ER α mRNA的显着多样性。(C)2009爱思唯尔有限公司保留所有权利。
The T-untranslated region (UTR) of the estrogen receptor alpha (ER alpha) gene plays an important role in determining tissue-specific expression. To elucidate the regulatory mechanisms of rat ERa gene expression, the genomic organization must be investigated. We therefore analyzed the structure of the rat ER alpha mRNA 5'-UTR using rapid amplification of 5'-cDNA ends (5'-RACE) and RT-PCR. The analysis showed the presence of multiple variants containing unique 5'-UTRs. We mapped the cDNA sequences on the rat genome, and newly identified one leader exon (exon 0U) and ten untranslated internal exons (exons 11-10). Both splicing from four different leader exons (exons 0S, 0N, 0U, and 0/B) onto exon 1 and alternative splicing in combination with eleven internal exons (exons I1-10, and OT) produce multiple transcripts. RT-PCR analysis revealed that each variant had preferred expression sites, suggesting that promoter usage and splicing are regulated in tissue-specific manners. Moreover, we determined a splicing event to yield Delta exon 1 variants (0S-2-3-4-5-6-7-8), which are translated into rat 46 kDa ER alpha proteins. Our results indicate that the rat ER alpha gene is more complex than previously thought in terms of genomic organization and that both alternative promoter usage and alternative splicing contribute to the remarkable diversity of ERa mRNAs. (C) 2009 Elsevier Ltd. All rights reserved.