Xu Y x, Ben-Shlomo H, Michaeli S
Department of Membrane Research and Biophysics, The Weizmann Institute of Science, Rehovot 76100, Israel.
Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8473-8. doi: 10.1073/pnas.94.16.8473.
Fractionation of the abundant small ribonucleoproteins (RNPs) of the trypanosomatid Leptomonas collosoma revealed the existence of a group of unidentified small RNPs that were shown to fractionate differently than the well-characterized trans-spliceosomal RNPs. One of these RNAs, an 80-nt RNA, did not possess a trimethylguanosine (TMG) cap structure but did possess a 5' phosphate terminus and an invariant consensus U5 snRNA loop 1. The gene coding for the RNA was cloned, and the coding region showed 55% sequence identity to the recently described U5 homologue of Trypanosoma brucei [Dungan, J. D., Watkins, K. P. & Agabian, N. (1996) EMBO J. 15, 4016-4029]. The L. collosoma U5 homologue exists in multiple forms of RNP complexes, a 10S monoparticle, and two subgroups of 18S particles that either contain or lack the U4 and U6 small nuclear RNAs, suggesting the existence of a U4/U6.U5 tri-small nuclear RNP complex. In contrast to T. brucei U5 RNA (62 nt), the L. collosoma homologue is longer (80 nt) and possesses a second stem-loop. Like the trypanosome U3, U6, and 7SL RNA genes, a tRNA gene coding for tRNACys was found 98 nt upstream to the U5 gene. A potential for base pair interaction between U5 and SL RNA in the 5' splice site region (positions -1 and +1) and downstream from it is proposed. The presence of a U5-like RNA in trypanosomes suggests that the most essential small nuclear RNPs are ubiquitous for both cis- and trans-splicing, yet even among the trypanosomatids the U5 RNA is highly divergent.
对锥虫科的粗线细滴虫丰富的小核糖核蛋白(RNP)进行分级分离,发现存在一组未鉴定的小RNP,其分级分离方式与特征明确的反式剪接体RNP不同。其中一种RNA,即80个核苷酸的RNA,不具有三甲基鸟苷(TMG)帽结构,但具有5'磷酸末端和不变的共有U5小核RNA环1。编码该RNA的基因被克隆,编码区与最近描述的布氏锥虫U5同源物显示出55%的序列同一性[邓根,J.D.,沃特金斯,K.P.和阿加比安,N.(1996年)《欧洲分子生物学组织杂志》15,4016 - 4029]。粗线细滴虫U5同源物以多种形式的RNP复合物存在,一种10S单颗粒,以及两个18S颗粒亚组,它们要么含有要么缺少U4和U6小核RNA,这表明存在U4/U6.U5三小核RNP复合物。与布氏锥虫U5 RNA(62个核苷酸)不同,粗线细滴虫同源物更长(80个核苷酸)且具有第二个茎环。与锥虫的U3、U6和7SL RNA基因一样,在U5基因上游98个核苷酸处发现了一个编码tRNACys的tRNA基因。有人提出在5'剪接位点区域(位置 -1和 +1)及其下游,U5和SL RNA之间存在碱基对相互作用的可能性。锥虫中存在U5样RNA表明,最基本的小核RNP对于顺式和反式剪接都是普遍存在的,然而即使在锥虫科中,U5 RNA也高度分化。