Inhibition of c-erbA mRNA splicing by a naturally occurring antisense RNA.
Inhibition of c-erbA mRNA splicing by a naturally occurring antisense RNA.
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DOI：
10.1016/s0021-9258(18)54535-x
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发表时间：
1991-11
期刊：
影响因子：
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通讯作者：
S. H. Munroe;Mitchell A. Lazar
中科院分区：
文献类型：
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作者：
S. H. Munroe;Mitchell A. Lazar
大鼠Erbaα基因座编码两个重叠的mRNAs，α1和α2，除了它们的最大3‘外显子外，它们是相同的。α1基因编码一种甲状腺激素受体，而α2基因编码一个功能未知的改变的配基结合域。先前的研究表明，在表达高水平的第三种RNA-Rev-ErbaαmRNA的细胞中，α1/α2的比率最高，该RNA以相反的方向转录，与α2互补，而不是α1 mRNA。假设碱基配对与REV-Erbaα阻止α2 mRNA的剪接，从而有利于非重叠α1的形成。为了验证这一模型，建立了一个在体外准确剪接α2前-mRNAs的系统。加入超过5倍的含有Rev-ErbaαmRNA 3‘端的反义RNA可抑制剪接。延伸到3‘剪接点的反义RNA和仅与外显子序列互补的较短的RNA都有效地阻止了剪接。然而，剪接只受到互补RNA的抑制。这些观察结果与碱基配对与互补RNA调节α1和α2 mRNAs的替代加工的机制是一致的。
The rat erbA alpha locus encodes two overlapping mRNAs, alpha 1 and alpha 2, which are identical except for their most 3' exons. alpha 1 mRNA encodes a thyroid hormone receptor, while alpha 2 encodes an altered ligand binding domain of unknown function. Previous studies have shown that the ratio of alpha 1 to alpha 2 is highest in cells expressing a high level of a third RNA, Rev-ErbA alpha mRNA, which is transcribed in the opposite direction and is complementary to alpha 2 but not alpha 1 mRNA. It was hypothesized that base pairing with Rev-ErbA alpha blocks splicing of alpha 2 mRNA, thereby favoring formation of the non-overlapping alpha 1. To test this model, a system was developed in which alpha 2 pre-mRNAs were accurately spliced in vitro. Splicing was inhibited by the addition of a 5-fold excess of antisense RNAs containing the 3' end of Rev-ErbA alpha mRNA. Both an antisense RNA extending across the 3' splice site and a shorter RNA complementary only to exon sequences efficiently blocked splicing. However, splicing was only inhibited by complementary RNAs. These observations are consistent with a mechanism in which base pairing with a complementary RNA regulates alternative processing of alpha 1 and alpha 2 mRNAs.