Partial purification of the yeast U2 snRNP reveals a novel yeast pre-mRNA splicing factor required for pre-spliceosome assembly
Partial purification of the yeast U2 snRNP reveals a novel yeast pre-mRNA splicing factor required for pre-spliceosome assembly
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DOI：
10.1093/emboj/18.12.3463
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发表时间：
1999-06-15
期刊：
影响因子：
11.4
通讯作者：
Séraphin, B
中科院分区：
文献类型：
--
作者：
Caspary, F;Shevchenko, A;Séraphin, B
我们已经部分纯化了酿酒酵母的 U2 snRNP。通过纳电喷雾质谱法对纯化组分中一致发现的一些蛋白质进行鉴定，表明存在一种名为 Rse1p 的新型剪接因子。 RSE1 基因是必需的，编码 148.2 kDa 的蛋白质。我们证明了 Rse1p 在低盐浓度下与 U2 snRNA 特异性结合。此外，我们还表明 Rse1p 是前剪接体的一个组成部分。 Rse1p 的耗尽和条件突变体的分析表明，Rse1p 是体内有效剪接所必需的。体外 Rse1p 是形成前剪接体所必需的。数据库搜索显示，Rse1p 在人类中是保守的，它属于一个大的蛋白质家族，其中包括多聚腺苷酸化因子和 DNA 修复蛋白。 Rse1p 的特征表明其人类同源物可能是 SF3 剪接因子的一个亚基。
We have partially purified the U2 snRNP of Saccharomyces cerevisiae. Identification of some proteins consistently found in the purified fractions by nanoelectrospray mass spectrometry indicated the presence of a novel splicing factor named Rse1p. The RSE1 gene is essential and codes for a 148.2 kDa protein. We demonstrated that Rse1p associates specifically with U2 snRNA at low salt concentrations. In addition, we showed that Rse1p is a component of the prespliceosome. Depletion of Rse1p and analysis of a conditional mutant indicated that Rse1p was required for efficient splicing in vivo. In vitro Rse1p is required for the formation of pre-spliceosomes. Database searches revealed that Rse1p is conserved in humans and that it belongs to a large protein family that includes polyadenylation factors and DNA repair proteins. The characteristics of Rse1p suggest that its human homologue could be a subunit of the SF3 splicing factor.