Blazey D L, Burns R O
Proc Natl Acad Sci U S A. 1982 Aug;79(16):5011-5. doi: 10.1073/pnas.79.16.5011.
Polarity of Tn10 insertion mutations in the Salmonella typhimurium ilvGEDA operon depends on both the location and the orientation of the Tn10 element. One orientation of Tn10 insertions in ilvG and ilvE permits low-level expression of the downstream ilvEDA and ilvDA genes, respectively. Our analysis of Salmonella ilv recombinant plasmids shows that this residual ilv expression must result from Tn10-directed transcription and does not reflect the presence of internal promoters in the ilvGEDA operon, as was previously suggested. The opposite orientation of Tn10 insertion in ilvE prevents ilvDA expression, indicating that only one end of Tn10 is normally active in transcribing adjacent genes. Both orientations of Tn10 insertion in ilvD exert absolute polarity on ilvA expression. Expression of ilvA is known to be dependent on effective translation of ilvD, perhaps reflecting the lack of a ribosome binding site proximal to the ilvA sequence. Therefore, recognition of the ability of Tn10 to promote transcription of contiguous genes in the ilvGEDA operon apparently requires the presence of associated ribosome binding sites.
鼠伤寒沙门氏菌ilvGEDA操纵子中Tn10插入突变的极性取决于Tn10元件的位置和方向。ilvG和ilvE中Tn10插入的一种方向分别允许下游的ilvEDA和ilvDA基因进行低水平表达。我们对沙门氏菌ilv重组质粒的分析表明,这种残留的ilv表达必定源于Tn10指导的转录,而并不像之前所认为的那样反映ilvGEDA操纵子中存在内部启动子。ilvE中Tn10插入的相反方向会阻止ilvDA表达,这表明通常只有Tn10的一端在转录相邻基因时具有活性。ilvD中Tn10插入的两个方向对ilvA表达均产生绝对极性。已知ilvA的表达依赖于ilvD的有效翻译,这可能反映出ilvA序列近端缺乏核糖体结合位点。因此,要认识到Tn10促进ilvGEDA操纵子中相邻基因转录的能力,显然需要存在相关的核糖体结合位点。