MacMorris Margaret, Kumar Madhur, Lasda Erika, Larsen Alison, Kraemer Brian, Blumenthal Thomas
Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Aurora, CO 80045, USA.
RNA. 2007 Apr;13(4):511-20. doi: 10.1261/rna.426707. Epub 2007 Feb 5.
In many Caenorhabditis elegans pre-mRNAs, the RNA sequence between the 5' cap and the first 3' splice site is replaced by trans-splicing a short spliced leader (SL) from the Sm snRNP, SL1. C. elegans also utilizes a similar Sm snRNP, SL2, to trans-splice at sites between genes in polycistronic pre-mRNAs from operons. How do SL1 and SL2 snRNPs function in different contexts? Here we show that the SL1 snRNP contains a complex of SL75p and SL21p, which are homologs of novel proteins previously reported in the Ascaris SL snRNP. Interestingly, we show that the SL2 snRNP does not contain these proteins. However, SL75p and SL26p, a paralog of SL21p, are components of another Sm snRNP that contains a novel snRNA species, Sm Y. Knockdown of SL75p is lethal. However, knockdown of either SL21p or SL26p alone leads to cold-sensitive sterility, whereas knockdown of both SL21p and SL26p is lethal. This suggests that these two proteins have overlapping functions even though they are associated with different classes of snRNP. These phenotypic relationships, along with the association of SL26p with SL75p, imply that, like the SL1 RNA/Sm/SL75p/SL21p complex, the Sm Y/Sm/SL75p/SL26p complex is associated with trans-splicing.
在许多秀丽隐杆线虫的前体信使核糖核酸(pre-mRNA)中,5' 帽与第一个3' 剪接位点之间的RNA序列会通过从Sm核小核糖核蛋白(snRNP)SL1转剪接一个短的剪接前导序列(SL)而被替换。秀丽隐杆线虫还利用一种类似的Sm snRNP,即SL2,在来自操纵子的多顺反子前体信使核糖核酸中基因之间的位点进行转剪接。SL1和SL2 snRNP在不同背景下是如何发挥作用的呢?在这里我们表明,SL1 snRNP包含SL75p和SL21p的复合物,它们是先前在蛔虫SL snRNP中报道的新型蛋白质的同源物。有趣的是,我们表明SL2 snRNP不包含这些蛋白质。然而,SL75p和SL21p的旁系同源物SL26p是另一种Sm snRNP的组成成分,该Sm snRNP包含一种新型的小核RNA(snRNA),即Sm Y。敲低SL75p是致死的。然而,单独敲低SL21p或SL26p会导致冷敏感不育,而同时敲低SL21p和SL26p则是致死的。这表明这两种蛋白质尽管与不同类别的snRNP相关,但具有重叠的功能。这些表型关系,连同SL26p与SL75p的关联,意味着,与SL1 RNA/Sm/SL75p/SL21p复合物一样,Sm Y/Sm/SL75p/SL26p复合物与转剪接相关。