Gray W J, Midgley J E
Biochem J. 1972 Aug;128(5):1007-20. doi: 10.1042/bj1281007.
The biosynthesis and stability of various RNA fractions was studied in RC(str) and RC(rel) multiple amino acid auxotrophs of Escherichia coli. In conditions of amino acid deprivation, RC(str) mutants were labelled with exogenous nucleotide bases at less than 1% of the rate found in cultures growing normally in supplemented media. Studies by DNA-RNA hybridization and by other methods showed that, during a period of amino acid withdrawal, not more than 60-70% of the labelled RNA formed in RC(str) mutants had the characteristics of mRNA. Evidence was obtained for some degradation of newly formed 16S and 23S rRNA species to heterogeneous material of lower molecular weight. This led to overestimations of the mRNA content of rapidly labelled RNA from such methods as simple examination of sucrose-density-gradient profiles. In RC(rel) strains the absolute and relative rates of synthesis of the various RNA fractions were not greatly affected. However, the stability of about half of the mRNA fraction was increased in RC(rel) strains during amino acid starvation, giving kinetics of mRNA labelling and turnover that were identical with those found in either RC(str) or RC(rel) strains inhibited by high concentrations of chloramphenicol. Coincidence hybridization techniques showed that the mRNA content of amino acid-starved RC(str) auxotrophs was unchanged from that found in normally growing cells. In contrast, RC(rel) strains deprived of amino acids increased their mRNA content about threefold. In such cultures the mRNA content of accumulating newly formed RNA was a constant 16% by wt.
在大肠杆菌的RC(str)和RC(rel)多种氨基酸营养缺陷型中研究了各种RNA组分的生物合成和稳定性。在氨基酸缺乏的条件下,RC(str)突变体用外源核苷酸碱基标记的速率低于在补充培养基中正常生长的培养物中发现的速率的1%。通过DNA-RNA杂交和其他方法进行的研究表明,在氨基酸撤出期间,RC(str)突变体中形成的标记RNA中不超过60-70%具有mRNA的特征。有证据表明新形成的16S和23S rRNA物种会降解为低分子量的异质物质。这导致通过简单检查蔗糖密度梯度图谱等方法高估了快速标记RNA的mRNA含量。在RC(rel)菌株中,各种RNA组分的绝对合成速率和相对合成速率没有受到很大影响。然而,在氨基酸饥饿期间,RC(rel)菌株中约一半的mRNA组分的稳定性增加,其mRNA标记和周转动力学与在高浓度氯霉素抑制的RC(str)或RC(rel)菌株中发现的相同。巧合杂交技术表明,氨基酸饥饿的RC(str)营养缺陷型的mRNA含量与正常生长细胞中的mRNA含量没有变化。相比之下,缺乏氨基酸的RC(rel)菌株的mRNA含量增加了约三倍。在这样的培养物中,积累的新形成RNA的mRNA含量按重量计恒定为16%。