Sipley J, Dunn J, Goldman E
Department of Microbiology and Molecular Genetics, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark 07103.
Mol Gen Genet. 1991 Dec;230(3):376-84. doi: 10.1007/BF00280294.
Bacteriophage T7 infection has been studied in Escherichia coli strains showing both increased and decreased ribosome fidelity and in the presence of streptomycin, which stimulates translational misreading, in an effort to determine effects on the apparent programmed translational frameshift that occurs during synthesis of the gene 10 capsid protein. Quantitation of the protein bands from SDS-PAGE failed to detect any significant effects on the amounts of the shifted 10B protein relative to the in-frame 10A protein under all fidelity conditions tested. However, any changes in fidelity conditions led to inhibition of phage morphogenesis in single-step growth experiments, which could not be accounted for by reduced amounts of phage protein synthesis, nor, at least in the case of decreased accuracy, by reduced amounts of phage DNA synthesis. Reduction in phage DNA synthesis did appear to account for a substantial proportion of the reduction in phage yield seen under conditions of increased accuracy. Similar effects of varying ribosomal fidelity on growth were also seen with phage T3, and to a lesser extent with phage T4. The absence of change in the high-frequency T7 gene 10 frameshift differs from earlier reports that ribosomal fidelity affects low-frequency frameshift errors.
为了确定对基因10衣壳蛋白合成过程中发生的明显程序性翻译移码的影响,研究了噬菌体T7在核糖体保真度增加和降低的大肠杆菌菌株中的感染情况,以及在存在链霉素(可刺激翻译错读)的情况下的感染情况。在所有测试的保真度条件下,SDS-PAGE蛋白条带的定量分析未能检测到相对于框内10A蛋白,移位的10B蛋白量有任何显著影响。然而,在单步生长实验中,保真度条件的任何变化都会导致噬菌体形态发生的抑制,这既不能用噬菌体蛋白质合成量的减少来解释,至少在准确性降低的情况下,也不能用噬菌体DNA合成量的减少来解释。噬菌体DNA合成的减少似乎确实是准确性提高条件下噬菌体产量降低的很大一部分原因。噬菌体T3也观察到核糖体保真度变化对生长有类似影响,噬菌体T4的影响较小。高频T7基因10移码没有变化,这与早期关于核糖体保真度影响低频移码错误的报道不同。