Melançon P, Leclerc D, Brakier-Gingras L
Département de Biochimie, Université de Montréal, Québec, Canada.
Biochim Biophys Acta. 1990 Aug 27;1050(1-3):98-103. doi: 10.1016/0167-4781(90)90148-u.
A deletion of five nucleotides was introduced at the 5' end of the Escherichia coli 16S rRNA gene cloned in an appropriate vector under control of a T7 promoter. The 16S rRNA generated by in vitro transcription could be assembled into 30S subunits. The deletion did not affect the efficiency of translation of natural messengers and the correct selection of the reading frame. However, it reduced the binding of the messengers, which suggests that the 5' end of 16S rRNA is located on the pathway followed by the messengers on the 30S subunits. The deletion also restricted the stimulation of misreading by streptomycin in a poly(U)-directed system. This is in accord with the proximity of the 5' end of 16S rRNA to proteins S4, S5 and S12, which are known to be involved in the control of translational accuracy.
在克隆于合适载体中并受T7启动子控制的大肠杆菌16S rRNA基因的5'端引入了五个核苷酸的缺失。通过体外转录产生的16S rRNA可组装成30S亚基。该缺失不影响天然信使的翻译效率和阅读框的正确选择。然而,它降低了信使的结合,这表明16S rRNA的5'端位于信使在30S亚基上所遵循的途径上。该缺失还限制了链霉素在聚(U)指导系统中对错读的刺激。这与16S rRNA的5'端靠近已知参与翻译准确性控制的蛋白质S4、S5和S12相一致。