Hanke P D, Fuchs J A
J Bacteriol. 1983 Jun;154(3):1040-5. doi: 10.1128/jb.154.3.1040-1045.1983.
A RNA-DNA hybridization assay for ribonucleoside diphosphate reductase (RDP reductase) mRNA was used to determine whether control of RDP reductase synthesis in Escherichia coli is at the level of RNA transcription. The correlation observed between the level of RDP reductase enzymatic activity and the rate of RDP reductase mRNA synthesis suggested that the control is at the level of RNA transcription. No increase in RDP reductase enzymatic activity or RDP reductase mRNA was observed during the first 15 min after removal of thymine from a thymine-requiring culture. Thereafter, the rate of RDP reductase mRNA synthesis increased linearly for approximately 75 min before beginning to level off. The addition of thymine to a culture starved for thymine resulted in a decreasing rate of RDP reductase mRNA synthesis. However, 30 min of growth in the presence of thymine was required before the rate of RDP reductase mRNA synthesis dropped to the rate observed in an exponentially growing culture. Inhibition of DNA synthesis caused by shifting a culture of a polC mutant or a dnaB mutant to nonpermissive growth conditions resulted in an increase in the rate of RDP reductase mRNA synthesis similar to that observed for a thymine-starved culture.
采用一种针对核糖核苷二磷酸还原酶(RDP还原酶)mRNA的RNA-DNA杂交试验,来确定大肠杆菌中RDP还原酶合成的调控是否发生在RNA转录水平。观察到的RDP还原酶酶活性水平与RDP还原酶mRNA合成速率之间的相关性表明,这种调控发生在RNA转录水平。在从需要胸腺嘧啶的培养物中去除胸腺嘧啶后的最初15分钟内,未观察到RDP还原酶酶活性或RDP还原酶mRNA增加。此后,RDP还原酶mRNA合成速率线性增加约75分钟,然后才开始趋于平稳。向胸腺嘧啶饥饿的培养物中添加胸腺嘧啶,导致RDP还原酶mRNA合成速率下降。然而,在胸腺嘧啶存在的情况下生长30分钟后,RDP还原酶mRNA合成速率才降至指数生长培养物中观察到的速率。将polC突变体或dnaB突变体的培养物转移到非允许生长条件下导致的DNA合成抑制,使得RDP还原酶mRNA合成速率增加,这与胸腺嘧啶饥饿培养物中观察到的情况相似。