Colombo P, Fried M
Eukaryotic Gene Organization and Expression Laboratory, Imperial Cancer Research Fund, London, UK.
Nucleic Acids Res. 1992 Jul 11;20(13):3367-73. doi: 10.1093/nar/20.13.3367.
The transcriptional initiation sites of the chicken ribosomal protein L7a (rpL7a) gene have been determined and found to occur at three consecutive cytidine residues at the start of a polypyrimidine tract of 8 base pairs (bp). A comparative analysis of the 5' upstream regions of the mouse, human and chicken rpL7a genes identified two sequence elements (Box A and Box B) conserved over the 600 million years of divergent evolution that separate mammals and birds. Only Box A (nts - 56 to - 39) and Box B (nts - 25 to - 4) sequences were detected to bind nuclear factors from mouse nuclear extracts in an analysis of the mouse rpL7a 5' upstream sequence. Box A and Box B bind different nuclear factors and the factor binding to mouse Box A and mouse Box B sequences could be effectively competed by corresponding homologous sequences from the human and chicken rpL7a promoters. These results indicate that elements of the rpL7a promoter region are conserved between mammals and birds. An in vivo analysis of the mouse rpL7a 5' upstream sequence required for efficient transcription identified the 5' border of the minimal promoter region as lying between nts - 50 and - 56. Constructs containing 56 bp of 5' upstream DNA and the first 25 bp rpL7a exon were very efficiently transcribed indicating that sequences within the first intron are not required for gene expression. No sequence similarity was detected between the rpL7a promoter elements and described promoter elements of other eukaryotic ribosomal protein genes.
鸡核糖体蛋白L7a(rpL7a)基因的转录起始位点已被确定,发现其位于一个8个碱基对(bp)的多嘧啶序列起始处的三个连续胞嘧啶残基处。对小鼠、人类和鸡rpL7a基因5'上游区域的比较分析确定了两个在哺乳动物和鸟类分化进化的6亿年中保守的序列元件(A框和B框)。在对小鼠rpL7a 5'上游序列的分析中,仅检测到A框(核苷酸 - 56至 - 39)和B框(核苷酸 - 25至 - 4)序列能与小鼠核提取物中的核因子结合。A框和B框结合不同的核因子,并且与小鼠A框和小鼠B框序列结合的因子能被人类和鸡rpL7a启动子的相应同源序列有效竞争。这些结果表明rpL7a启动子区域的元件在哺乳动物和鸟类之间是保守的。对高效转录所需的小鼠rpL7a 5'上游序列的体内分析确定最小启动子区域的5'边界位于核苷酸 - 50至 - 56之间。包含5'上游DNA的56 bp和rpL7a外显子的前25 bp的构建体被高效转录,这表明基因表达不需要第一个内含子内的序列。在rpL7a启动子元件与其他真核核糖体蛋白基因的所述启动子元件之间未检测到序列相似性。