Schimanski Bernd, Laufer Gabriele, Gontcharova Lilia, Günzl Arthur
Center for Microbial Pathogenesis, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030-3710, USA.
Nucleic Acids Res. 2004 Feb 2;32(2):700-9. doi: 10.1093/nar/gkh231. Print 2004.
In the protist parasite Trypanosoma brucei, the small nuclear spliced leader (SL) RNA and the large rRNAs are key molecules for mRNA maturation and protein synthesis, respectively. The SL RNA gene (SLRNA) promoter recruits RNA polymerase II and consists of a bipartite upstream sequence element (USE) and an element close to the transcription initiation site. Here, we analyzed the distal part of the ribosomal (RRNA) promoter and identified two sequence blocks which, in reverse orientation, closely resemble the SLRNA USE by both sequence and spacing. A detailed mutational analysis revealed that the ribosomal (r)USE is essential for efficient RRNA transcription in vivo and that it functions in an orientation-dependent manner. Moreover, we showed that USE and rUSE are functionally interchangeable and that rUSE stably interacted with an essential factor of SLRNA transcription. Finally, we demonstrated that the T.brucei homolog of the recently characterized transcription factor p57 of the related organism Leptomonas seymouri specifically bound to USE and rUSE. Since p57 and its T.brucei counterpart are homologous to SNAP50, a component of the human small nuclear RNA gene activation protein complex (SNAPc), both SLRNA and RRNA transcription in T.brucei may depend on a SNAPc-like transcription factor.
在原生生物寄生虫布氏锥虫中,小核剪接前导(SL)RNA和大核糖体RNA(rRNA)分别是mRNA成熟和蛋白质合成的关键分子。SL RNA基因(SLRNA)启动子招募RNA聚合酶II,由一个二分体上游序列元件(USE)和一个靠近转录起始位点的元件组成。在这里,我们分析了核糖体(RRNA)启动子的远端部分,并鉴定出两个序列块,它们以反向方向在序列和间距上都与SLRNA USE非常相似。详细的突变分析表明,核糖体(r)USE对于体内高效的RRNA转录至关重要,并且它以方向依赖的方式发挥作用。此外,我们表明USE和rUSE在功能上是可互换的,并且rUSE与SLRNA转录的一个必需因子稳定相互作用。最后,我们证明了相关生物西氏细滴虫最近鉴定的转录因子p57的布氏锥虫同源物特异性结合USE和rUSE。由于p57及其布氏锥虫对应物与人类小核RNA基因激活蛋白复合物(SNAPc)的一个组分SNAP50同源,布氏锥虫中的SLRNA和RRNA转录可能都依赖于一种类似SNAPc的转录因子。