Fayat G, Mayaux J F, Sacerdot C, Fromant M, Springer M, Grunberg-Manago M, Blanquet S
J Mol Biol. 1983 Dec 15;171(3):239-61. doi: 10.1016/0022-2836(83)90092-x.
The nucleotide sequences of pheS and of the beginning of pheT have been determined. The genes pheS and pheT code, respectively, for the small and large subunits of phenylalanyl-tRNA synthetase, an alpha 2 beta 2 enzyme. Upstream from pheS the sequence shows another open reading frame of 354 nucleotides (rplT), which accounts for a protein of Mr 13,400. The product of this gene, previously named "P12", is identified as the ribosomal protein L20. The promoter for the pheS, T operon was located 368 nucleotides in front of pheS by transcription experiments in vitro. The promoter site is followed by a short open reading frame, which codes for a 14-residue peptide containing five phenylalanine residues. Immediately downstream from the stop codon of this open reading frame, the DNA sequence indicates that the transcript can be folded into three alternative secondary structures, one of which is a site of transcription termination. In vitro, 90% of transcription products initiated at the pheS, T promoter terminate at this site. However, long run-off transcripts proceeding through the terminator and covering the pheS structural gene are observed. No other transcription initiation could be detected between the terminator and the pheS structural gene. All these results are consistent with a mechanism by which phenylalanine-mediated attenuation controls the expression of phenylalanyl-tRNA synthetase. Further evidence is provided for this model by the features of pheS, T regulation in vivo (see the accompanying paper).
已确定了pheS基因的核苷酸序列以及pheT基因起始部分的核苷酸序列。基因pheS和pheT分别编码苯丙氨酰 - tRNA合成酶的小亚基和大亚基,该酶是一种α2β2型酶。在pheS上游,该序列显示出另一个由354个核苷酸组成的开放阅读框(rplT),它编码一个分子量为13400的蛋白质。这个基因的产物,先前被命名为“P12”,被鉴定为核糖体蛋白L20。通过体外转录实验,确定pheS、T操纵子的启动子位于pheS前方368个核苷酸处。启动子位点之后是一个短的开放阅读框,它编码一个含有五个苯丙氨酸残基的14个氨基酸残基的肽段。在这个开放阅读框的终止密码子紧下游,DNA序列表明转录本可以折叠成三种不同的二级结构,其中之一是转录终止位点。在体外,从pheS、T启动子起始的转录产物中90%在这个位点终止。然而,也观察到有长的溢流转录本穿过终止子并覆盖pheS结构基因。在终止子和pheS结构基因之间未检测到其他转录起始。所有这些结果都与苯丙氨酸介导的衰减控制苯丙氨酰 - tRNA合成酶表达的机制相一致。体内pheS、T调控的特征为该模型提供了进一步的证据(见随附论文)。