Aiso Toshiko, Yoshida Hideji, Wada Akira, Ohki Reiko
Department of Molecular Biology, School of Health Sciences, Kyorin University, Hachioji, Tokyo, Japan.
J Bacteriol. 2005 Mar;187(6):1951-8. doi: 10.1128/JB.187.6.1951-1958.2005.
The expression of ribosome modulation factor (RMF) is induced during stationary phase in Escherichia coli. RMF participates in the dimerization of 70S ribosomes to form the 100S ribosome, which is the translationally inactive form of the ribosome. To elucidate the involvement of the control of mRNA stability in growth-phase-specific rmf expression, we investigated rmf mRNA stability in stationary-phase cells and cells inoculated into fresh medium. The rmf mRNA was found to have an extremely long half-life during stationary phase, whereas destabilization of this mRNA took place after the culture was inoculated into fresh medium. RMF and 100S ribosomes disappeared from cells 1 min after inoculation. In addition to control by ppGpp-dependent transcription, these results indicate that the modulation of rmf mRNA stability is also involved in the regulation of growth-phase-specific rmf expression. Unexpectedly, the postinoculation degradation of rmf mRNA was suppressed by the addition of rifampin, suggesting that de novo RNA synthesis is necessary for degradation. This degradation was also suppressed in both a poly(A) polymerase-deficient and an rne-131 mutant strain. We cloned and sequenced the 3'-proximal regions of rmf mRNAs and found that most of these 3' ends terminated at the rho-independent terminator with the addition of a one- to five-A oligo(A) tail in either stationary-phase or inoculated cells. No difference was observed in the length of the poly(A) tail between stationary-phase and inoculated cells. These results suggest that a certain postinoculation-specific regulatory factor participates in the destabilization of rmf mRNA and is dependent on polyadenylation.
核糖体调控因子(RMF)的表达在大肠杆菌的稳定期被诱导。RMF参与70S核糖体的二聚化以形成100S核糖体,后者是核糖体的翻译无活性形式。为了阐明mRNA稳定性控制在生长阶段特异性rmf表达中的作用,我们研究了稳定期细胞和接种到新鲜培养基中的细胞中rmf mRNA的稳定性。发现rmf mRNA在稳定期具有极长的半衰期,而将培养物接种到新鲜培养基后,该mRNA会发生去稳定化。接种后1分钟,RMF和100S核糖体从细胞中消失。除了受ppGpp依赖的转录控制外,这些结果表明rmf mRNA稳定性的调节也参与了生长阶段特异性rmf表达的调控。出乎意料的是,接种后rmf mRNA的降解被利福平的添加所抑制,这表明从头RNA合成对于降解是必需的。在聚腺苷酸聚合酶缺陷型和rne - 131突变株中,这种降解也受到抑制。我们克隆并测序了rmf mRNA的3'近端区域,发现大多数这些3'末端在稳定期或接种细胞中都在不依赖ρ的终止子处终止,并添加了一到五个A的寡聚(A)尾巴。在稳定期细胞和接种细胞之间,聚(A)尾巴的长度没有差异。这些结果表明,某种接种后特异性调节因子参与了rmf mRNA的去稳定化,并且依赖于多聚腺苷酸化。