Georgellis D, Arvidson S, von Gabain A
Karolinska Institute, Department of Bacteriology, Stockholm, Sweden.
J Bacteriol. 1992 Aug;174(16):5382-90. doi: 10.1128/jb.174.16.5382-5390.1992.
By growing Escherichia coli in continuous cultures at various growth rates, we provide definitive evidence that the stability of the ompA mRNA is growth rate dependent. Shifting fast-growing cells into physiological salt buffer led to an immediately increased rate of ompA mRNA decay and to an instantly decreased rate of 9S RNA conversion into 5S rRNA. Shifting slowly growing cells into fresh medium had the opposite effect for each of the two RNA species. The observed regulatory patterns underline the need of cells to adjust the output of ompA and 9S RNAs in response to growth rate changes. At all growth rates and throughout all shift experiments, the half-life of bla mRNA was constant. A stabilization of the ompA transcript was even observed when slowly growing cells were shifted into fresh medium already containing the transcriptional inhibitor rifampicin. A hybrid bla transcript with the 5' untranslated region from the ompA gene behaved similarly to the wild-type ompA messenger in response to a shift in growth rate. In agreement with this result, we found that the same type of 5' cleavages as have been previously shown to initiate the decay of the ompA transcript seem to be involved in stability regulation. In E. coli the degradation of mRNA has been shown to depend on the ams/rne gene. This gene controls the stability-related cleavages in the ompA transcript, catabolic processes, and the cleavages which process the 9S rRNA into 5S RNA, an anabolic process. We discuss these results with respect to the ams/rne gene and the related nuclease activities that control the ompA and 9S RNA cleavages.
通过在不同生长速率下对大肠杆菌进行连续培养,我们提供了确凿证据,证明ompA mRNA的稳定性取决于生长速率。将快速生长的细胞转移到生理盐缓冲液中会导致ompA mRNA的衰变速率立即增加,以及9S RNA转化为5S rRNA的速率立即降低。将缓慢生长的细胞转移到新鲜培养基中对这两种RNA的影响则相反。观察到的调控模式突显了细胞根据生长速率变化调整ompA和9S RNA输出的必要性。在所有生长速率下以及在所有转移实验中,bla mRNA的半衰期都是恒定的。当缓慢生长的细胞转移到已经含有转录抑制剂利福平的新鲜培养基中时,甚至观察到ompA转录本的稳定化。一个带有ompA基因5'非翻译区的杂交bla转录本在生长速率发生变化时的表现与野生型ompA信使相似。与这一结果一致,我们发现,之前已证明能引发ompA转录本衰变的相同类型的5'切割似乎参与了稳定性调控。在大肠杆菌中,mRNA的降解已被证明依赖于ams/rne基因。该基因控制ompA转录本中与稳定性相关的切割、分解代谢过程,以及将9S rRNA加工成5S RNA的切割,后者是一个合成代谢过程。我们结合ams/rne基因以及控制ompA和9S RNA切割的相关核酸酶活性来讨论这些结果。