Functioning of the Drosophila Wilms'-tumor-1-associated protein homolog, Fl(2)d, in sex-lethal-dependent alternative splicing
Functioning of the Drosophila Wilms'-tumor-1-associated protein homolog, Fl(2)d, in sex-lethal-dependent alternative splicing
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DOI：
10.1534/genetics.107.081679
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发表时间：
2008-02-01
期刊：
影响因子：
3.3
通讯作者：
Schedl, Paul
中科院分区：
文献类型：
--
作者：
Penn, Jill K. M.;Graham, Patricia;Schedl, Paul
fl(2)d是Wilms'-tumor-1-associated protein （WTAP）的果蝇同源物，可调节性致死性（Sxl）、变性（tra）和超abithorax （Ubx）的选择性剪接。尽管WTAP已在功能性人类剪接体中被发现，但它究竟如何参与剪接过程仍不清楚。在这里，我们试图确定与fl(2)d在遗传和物理上相互作用的因素。我们首先详细分析了Sxl-Fl(2)d蛋白-蛋白相互作用，并提出证据表明雌性特异性fl(2)d等位基因在性别决定过程中是反同的。接下来，我们发现fl(2)d与早期作用的一般剪接调节因子相互作用，并且fl(2)d存在于Snf、U2AF50、U2AF38和U1-70K的免疫沉淀复合物中。相比之下，我们无法检测到含有U5 snRNP蛋白U5- 40k或与活化的B剪接体复合物SKIP相关的蛋白的fl(2)d复合物。值得注意的是，在Sxl中观察到的遗传和分子相互作用与fl(2)d中检测到的非常相似。综上所述，我们的研究结果表明Sxl和fl(2)d在剪接体组装的早期阶段改变了剪接位点的选择。
fl(2)d, the Drosophila homolog of Wilms'-tumor-1-associated protein (WTAP), regulates the alternative splicing of Sex-lethal (Sxl), transformer (tra), and Ultrabithorax (Ubx). Although WTAP has been found in functional human spliceosomes, exactly how it contributes to the splicing process remains unknown. Here we attempt to identify factors that interact genetically and physically with fl(2)d. We begin by analyzing the Sxl-Fl(2)d protein-protein interaction in detail and present evidence suggesting that the female-specific fl(2)d' allele is antimorphic with respect to the process of sex determination. Next we show that fl(2)d interacts genetically with early acting general splicing regulators and that Fl(2)d is present in immunoprecipitable complexes with Snf, U2AF50, U2AF38, and U1-70K By contrast, we could not detect Fl(2)d complexes containing the U5 snRNP protein U5-40K or with a protein that associates with the activated B spliceosomal complex SKIP. Significantly, the genetic and molecular interactions observed for Sxl are quite similar to those detected for fl(2)d. Taken together, our findings suggest that Sxl and fl(2)d function to alter splice-site selection at an early step in spliceosome assembly.