Leclerc D, Melançon P, Brakier-Gingras L
Département de Biochimie, Université de Montréal, Canada.
Nucleic Acids Res. 1991 Jul 25;19(14):3973-7. doi: 10.1093/nar/19.14.3973.
The nine possible single-base substitutions were produced at positions 913 to 915 of the 16S ribosomal RNA of Escherichia coli, a region known to be protected by streptomycin [Moazed, D. and Noller, H.F. (1987) Nature, 327, 389-394]. When the mutations were introduced into the expression vector pKK3535, only two of them (913A----G and 915A----G) permitted recovery of viable transformants. Ribosomes were isolated from the transformed bacteria and were assayed for their response to streptomycin in poly(U)- and MS2 RNA-directed assays. They were resistant to the stimulation of misreading and to the inhibition of protein synthesis by streptomycin, and this correlated with a decreased binding of the drug. These results therefore demonstrate that, in line with the footprinting studies of Moazed and Noller, mutations in the 915 region alter the interaction between the ribosome and streptomycin.
在大肠杆菌16S核糖体RNA的913至915位产生了9种可能的单碱基替换，该区域已知受链霉素保护[莫阿泽德，D.和诺勒，H.F.（1987年）《自然》，327，389 - 394]。当将这些突变引入表达载体pKK3535时，其中只有两个（913A→G和915A→G）能使活的转化体得以恢复。从转化后的细菌中分离出核糖体，并在聚（U）和MS2 RNA指导的试验中检测它们对链霉素的反应。它们对链霉素诱导的错读刺激和蛋白质合成抑制具有抗性，这与药物结合减少相关。因此，这些结果表明，与莫阿泽德和诺勒的足迹研究一致，915区域的突变改变了核糖体与链霉素之间的相互作用。
