Lee J Y, Evans C F, Engelke D R
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109-0606.
Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6986-90. doi: 10.1073/pnas.88.16.6986.
The RNA subunit of Saccharomyces cerevisiae nuclear RNase P is encoded by a single-copy, essential gene, RPR1. The 369-nucleotide mature form of the RNA has an apparent precursor with an 84-nucleotide 5' leader and approximately 33 nucleotides of additional 3' sequence. Analysis of RPR1 transcription in a strain with a temperature-sensitive lesion in RNA polymerase III shows that the gene is transcribed in vivo by RNA polymerase III. Examination of potential promoter regions using both progressive upstream deletions and point mutations indicates that at least two sequences contained within the 5' leader region are essential for expression in vivo, while sequences farther upstream influence efficiency. The required leader elements resemble tRNA gene-like A-box and B-box internal promoters in sequence and spacing. As in the tRNA genes, transcription factor TFIIIC binds to this region in vitro and binding is severely reduced by either A-box or B-box point mutations that impair expression in vivo. It thus appears that the yeast RNase P RNA gene has adopted a promoter strategy that places an RNA polymerase III "internal" promoter upstream of the mature structural domain to help drive transcription.
酿酒酵母核酶P的RNA亚基由单拷贝的必需基因RPR1编码。RNA的369个核苷酸的成熟形式有一个明显的前体,其5'端有一个84个核苷酸的前导序列和大约33个核苷酸的额外3'端序列。在RNA聚合酶III中存在温度敏感损伤的菌株中对RPR1转录进行分析,结果表明该基因在体内由RNA聚合酶III转录。使用渐进式上游缺失和点突变对潜在启动子区域进行检测,结果表明5'端前导序列中包含的至少两个序列对于体内表达至关重要,而更上游的序列影响表达效率。所需的前导元件在序列和间距上类似于tRNA基因样的A盒和B盒内部启动子。与tRNA基因一样,转录因子TFIIIC在体外与该区域结合,并且A盒或B盒点突变会严重降低结合,这些突变会损害体内表达。因此,酵母核糖核酸酶P RNA基因似乎采用了一种启动子策略,即将RNA聚合酶III“内部”启动子置于成熟结构域的上游以帮助驱动转录。