PRIMARY STRUCTURE OF THE HUMAN SPLICING FACTOR ASF REVEALS SIMILARITIES WITH DROSOPHILA REGULATORS
PRIMARY STRUCTURE OF THE HUMAN SPLICING FACTOR ASF REVEALS SIMILARITIES WITH DROSOPHILA REGULATORS
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DOI：
10.1016/0092-8674(91)90626-a
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发表时间：
1991-07-26
期刊：
影响因子：
64.5
通讯作者：
MANLEY, JL
中科院分区：
文献类型：
--
作者：
GE, H;ZUO, P;MANLEY, JL
我们之前描述了一种名为选择性剪接因子(ASF)的人类蛋白质的纯化，该蛋白质可以切换SV40早期Pre-mRNA中替代5‘剪接位点的利用。我们现在报告分离到一个编码该蛋白的cdna，命名为ASF-1。ASF-1由248个氨基酸残基组成，N-端有一个80个残基的RNA结合区，C-末端有一个50个残基，由80%的丝氨酸+精氨酸组成。在大肠杆菌中产生的ASF-1可以激活体外剪接，并切换5‘剪接位点的利用，证明重组蛋白足以提供这些活性。对额外的cDNA的分析表明，令人惊讶的是，ASF前-mRNA本身可以通过利用一个共享的5‘剪接位点和两个间隔很近的3’剪接位点进行交替剪接。使用上游位点导致第二个mRNA(ASF-2)，其中下游外显子的广泛翻译发生在与ASF-1不同的另一个阅读框架中。
We described previously the purification of a human protein, called alternative splicing factor (ASF), that can switch utilization of alternative 5' splice sites in an SV40 early pre-mRNA. We now report the isolation of a cDNA, designated ASF-1, that encodes this protein. ASF-1 consists of 248 amino acid residues, including an 80 residue RNA-binding domain at its N-terminus and a 50 residue C-terminal region that is 80% serine plus arginine. ASF-1 produced in E. coli can activate splicing in vitro and switch 5' splice-site utilization, establishing that the recombinant protein is sufficient to supply these activities. Analysis of additional cDNAs revealed that ASF pre-mRNA can itself be alternatively spliced, surprisingly, by utilization of a shared 5' splice site and two closely spaced 3' splice sites. Use of the upstream site results in a second mRNA (ASF-2) in which translation of the downstream exon occurs extensively in an alternative reading frame distinct from ASF-1.