Zorio D A, Blumenthal T
Department of Biology, Indiana University, Bloomington 47405, USA.
RNA. 1999 Apr;5(4):487-94. doi: 10.1017/s1355838299982225.
In most species the 3' splice site is recognized initially by an interaction between the two-subunit splicing factor U2AF with the polypyrimidine (poly(Y)) tract that results in recruitment of the U2 snRNP to the branch-point consensus just upstream. In contrast, in Caenorhabditis elegans, both the poly(Y) tract and the branch-point consensus sequences are missing, apparently replaced by the highly conserved U4CAG/R 3' splice site consensus. Nevertheless C. elegans U2AF65 is very similar to its mammalian and fly counterparts and may recognize the 3' splice site consensus. Here we report the cloning of the C. elegans U2AF35 gene, uaf-2. We show that it lacks an identifiable RS domain, which, in flies, has been shown to play a role in RNA binding, but it contains an extended glycine-rich stretch at its C-terminus. uaf-2 is in an operon with cyp-13, a gene that encodes a cyclophilin with an RRM domain at its N-terminus. We demonstrate by RNA interference that both U2AF genes, uaf-1 (which encodes U2AF65) and uaf-2, are required for viability, whereas cyp-13 is apparently not.
在大多数物种中,3' 剪接位点最初是通过两亚基剪接因子U2AF与多嘧啶(poly(Y))序列之间的相互作用来识别的,这种相互作用导致U2 snRNP被招募到上游的分支点共有序列处。相比之下,在秀丽隐杆线虫中,多嘧啶序列和分支点共有序列均缺失,显然被高度保守的U4CAG/R 3' 剪接位点共有序列所取代。然而,秀丽隐杆线虫的U2AF65与其哺乳动物和果蝇的对应物非常相似,可能识别3' 剪接位点共有序列。在此,我们报道了秀丽隐杆线虫U2AF35基因uaf-2的克隆。我们发现它缺乏可识别的RS结构域,在果蝇中,该结构域已被证明在RNA结合中起作用,但它在其C末端含有一个延伸的富含甘氨酸的区域。uaf-2与cyp-13处于同一个操纵子中,cyp-13是一个在其N末端编码具有RRM结构域的亲环蛋白的基因。我们通过RNA干扰证明,两个U2AF基因,即uaf-1(编码U2AF65)和uaf-2,对于生存力是必需的,而cyp-13显然不是。