Grondal E J, Evers R, Cornelissen A W
Max-Planck-Institut für Biologie, Molecular Parasitology Unit, Tübingen, FRG.
Nucleic Acids Res. 1990 Mar 25;18(6):1333-8. doi: 10.1093/nar/18.6.1333.
We have identified the promoter region of the large ribosomal DNA repeat unit of Crithidia fasciculata by northern blotting and nuclear run-on analyses. These data show that transcription starts approximately 1 kb upstream of the 18S rRNA gene. S1 protection experiments and sequence analysis of this area resulted in a precise localization of the start site. We have been unable to identify conserved sequence element(s) by a direct comparison of the crithidial RNA polymerase I promoter region and similar promoter regions of other eukaryotes; not even to the promoter region of the more closely related kinetoplastid species, Trypanosoma brucei. The absence of homology within the primary sequence of the promoter region, which is also found in other eukaryotes, might explain the observed species specificity of in vivo and in vitro rDNA transcription, since this resides in the interaction of initiation factor(s) and the core promoter domain.
我们通过Northern印迹法和细胞核连续转录分析确定了纤细短膜虫大核糖体DNA重复单元的启动子区域。这些数据表明转录起始于18S rRNA基因上游约1 kb处。对该区域进行的S1核酸酶保护实验和序列分析确定了起始位点的精确位置。通过直接比较短膜虫RNA聚合酶I启动子区域与其他真核生物的类似启动子区域,我们未能鉴定出保守的序列元件;甚至与亲缘关系更近的动基体物种布氏锥虫的启动子区域相比也未发现。启动子区域一级序列中缺乏同源性(其他真核生物中也存在这种情况),这可能解释了在体内和体外rDNA转录中观察到的物种特异性,因为这种特异性存在于起始因子与核心启动子结构域的相互作用中。