Hager P W, Rabinowitz J C
J Biol Chem. 1985 Dec 5;260(28):15163-7.
Bacillus subtilis 30 S subunits inefficiently recognize initiation sites in mRNAs from Gram-negative bacteria, but they are able to efficiently recognize initiation sites in mRNA derived from Gram-positive bacteria. McLaughlin et al. (McLaughlin, J. R., Murray, C. L., and Rabinowitz, J. C. (1981) J. Biol. Chem. 256, 11283-11291) have suggested that B. subtilis ribosomes require a strong Shine-Dalgarno sequence for translation initiation. To test whether this criterion is sufficient to explain the translational specificity of B. subtilis ribosomes, T7 late mRNA, which contains strong Shine-Dalgarno sequences before many of the late genes (Dunn, J. J., and Studier, F. W. (1983) J. Mol. Biol. 166, 477-535), was translated in vitro with both Escherichia coli and B. subtilis ribosomes. The identification of several of the in vitro products upon gel electrophoresis indicated that B. subtilis ribosomes recognize correct translation initiation sites in late T7 mRNA, but they do not translate these products efficiently. Competition experiments demonstrated that late T7 mRNA does not inhibit B. subtilis ribosomal translation of B. subtilis derived mRNA (from the bacteriophage phi 29). It is concluded that strong Shine-Dalgarno sequences may be necessary in B. subtilis translation initiation sites; however, additional determinants of initiation which differ from those found in the translation initiation sites of E. coli mRNAs must exist.
枯草芽孢杆菌的30 S亚基难以有效识别革兰氏阴性菌mRNA中的起始位点,但能够有效识别革兰氏阳性菌来源mRNA中的起始位点。麦克劳克林等人(麦克劳克林,J. R.,默里,C. L.,以及拉比诺维茨,J. C.(1981年)《生物化学杂志》256卷,11283 - 11291页)提出,枯草芽孢杆菌核糖体在翻译起始时需要一个强的夏因 - 达尔加诺序列。为了测试该标准是否足以解释枯草芽孢杆菌核糖体的翻译特异性,含有许多晚期基因之前具有强夏因 - 达尔加诺序列的T7晚期mRNA(邓恩,J. J.,以及斯图迪尔,F. W.(1983年)《分子生物学杂志》166卷,477 - 535页)在体外与大肠杆菌和枯草芽孢杆菌核糖体一起进行翻译。凝胶电泳后对几种体外产物的鉴定表明,枯草芽孢杆菌核糖体识别晚期T7 mRNA中的正确翻译起始位点,但它们不能有效地翻译这些产物。竞争实验表明,晚期T7 mRNA不会抑制枯草芽孢杆菌核糖体对枯草芽孢杆菌来源mRNA(来自噬菌体φ29)的翻译。得出的结论是,强夏因 - 达尔加诺序列在枯草芽孢杆菌翻译起始位点可能是必要的;然而,必须存在与大肠杆菌mRNA翻译起始位点中发现的决定因素不同的其他起始决定因素。