Shwed P S, Nazar R N
Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Nucleic Acids Res. 1999 Jul 15;27(14):2883-8. doi: 10.1093/nar/27.14.2883.
RNA polymerase I transcripts, purified from Schizosaccharomyces pombe cells, terminate at three sites that precede 'Sal box'-like termination element (TE) sequences. Essential features in these elements were investigated by the in vivo expression of targeted mutations. RNA analyses confirmed a functional significance for two of the elements (Boxes 1 and 3), but indicated that the third, less related, sequence (Box 2) does not function as a termination signal. The results further indicated that the most conserved residues in the two active TEs, as well as adjacent regions, are also most critical to function. Furthermore, some mutations in these elements or in immediately flanking sequences affect not only the efficiency of termination, but also alter the position of termination by as much as 35 nt. Since the element is able to influence the site of termination over a surprisingly long stretch of DNA sequence, these observations suggest that the TE does not act simply as a pause element by fixing the termination factor.
从粟酒裂殖酵母细胞中纯化得到的RNA聚合酶I转录本在三个位于类似“Sal盒”终止元件(TE)序列之前的位点终止。通过对靶向突变的体内表达研究了这些元件的基本特征。RNA分析证实了其中两个元件(盒1和盒3)具有功能意义,但表明第三个相关性较低的序列(盒2)不充当终止信号。结果进一步表明,两个活性TE中最保守的残基以及相邻区域对功能也最为关键。此外,这些元件或紧邻序列中的一些突变不仅影响终止效率,还会使终止位置改变多达35个核苷酸。由于该元件能够在一段惊人的长DNA序列上影响终止位点,这些观察结果表明TE并非简单地通过固定终止因子来充当暂停元件。