The serine/arginine-rich protein family in rice plays important roles in constitutive and alternative splicing of pre-mRNA
The serine/arginine-rich protein family in rice plays important roles in constitutive and alternative splicing of pre-mRNA
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DOI：
10.1105/tpc.105.037069
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发表时间：
2006-01-01
期刊：
影响因子：
11.6
通讯作者：
Shimamoto, K
中科院分区：
文献类型：
--
作者：
Isshiki, M;Tsumoto, A;Shimamoto, K
Ser/Arg-rich （SR）蛋白在pre-mRNA的组成剪接和选择性剪接中起重要作用。从水稻（Oryza sativa）中分离到20个编码SR蛋白的基因，其中6个基因具有植物特异性。为了确定SR蛋白是否调节水稻pre-mRNA的剪接效率和选择性剪接，我们通过将SR蛋白基因与水稻Waxy(b) (Wx(b))- β -葡萄糖醛酸酶融合基因共转化，在水稻原生质体中进行了瞬时测定。结果表明，植物特异性RSp29和与人类9G8同源的SR蛋白RSZp23增强了Wx(b)内含子1的剪接，并改变了Wx(b)内含子1的备选5'剪接位点。每个SR蛋白的剪接模式都是独一无二的；RSp29刺激远端剪接，而RSZp23促进近端剪接。植物特异性RSp29与sc26（人类SC35的同源物）之间的结构域交换实验结果表明，RNA识别基序1和Arg/Ser-rich （RS）结构域对提高剪接效率的重要性。在转基因水稻中，植物特异性RSZ36和SRp33b（人类ASF/SF2的同源物）的过表达改变了它们自身的前mrna和其他SR蛋白的选择性剪接模式。这些结果表明SR蛋白在水稻pre-mRNA的组成剪接和选择性剪接中起重要作用。
Ser/Arg-rich (SR) proteins play important roles in the constitutive and alternative splicing of pre-mRNA. We isolated 20 rice (Oryza sativa) genes encoding SR proteins, of which six contain plant-specific characteristics. To determine whether SR proteins modulate splicing efficiency and alternative splicing of pre-mRNA in rice, we used transient assays in rice protoplasts by cotransformation of SR protein genes with the rice Waxy(b) (Wx(b))-beta-glucuronidase fusion gene. The results showed that plant-specific RSp29 and RSZp23, an SR protein homologous to human 9G8, enhanced splicing and altered the alternative 5' splice sites of Wx(b) intron 1. The resulting splicing pattern was unique to each SR protein; RSp29 stimulated splicing at the distal site, and RSZp23 enhanced splicing at the proximal site. Results of domain-swapping experiments between plant-specific RSp29 and SCL26, which is a homolog of human SC35, showed the importance of RNA recognition motif 1 and the Arg/Ser-rich (RS) domain for the enhancement of splicing efficiencies. Overexpression of plant-specific RSZ36 and SRp33b, a homolog of human ASF/SF2, in transgenic rice changed the alternative splicing patterns of their own pre-mRNAs and those of other SR proteins. These results show that SR proteins play important roles in constitutive and alternative splicing of rice pre-mRNA.