Apirakaramwong A, Fukuchi J, Kashiwagi K, Kakinuma Y, Ito E, Ishihama A, Igarashi K
Faculty of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba, 263-8522, Japan.
Biochem Biophys Res Commun. 1998 Oct 20;251(2):482-7. doi: 10.1006/bbrc.1998.9494.
Ribosome modulation factor (RMF) is involved in stabilization of ribosomes during the transition from exponential growth to the stationary growth phase in Escherichia coli. A deficiency of RMF is known to reduce cell viability. Overaccumulation of spermidine also leads to a decrease in cell viability and to a decrease in the synthesis of RMF and of the cation-selective porin OmpC. Thus, a decrease in RMF levels may be involved in the decreased cell viability caused by excess spermidine. Because spermidine also influences the expression of OmpC, we examined whether OmpC deficiency enhances the cell death caused by RMF deficiency. The ompC mutant by itself did not affect protein synthesis or cell viability, but the double rmf ompC mutant produced a much larger decrease in protein synthesis and cell viability than did the single rmf mutant. There was also a decrease in the amount of ribosomes and in the Mg2+ content in the double rmf ompC mutant, and cell viability could be partially restored by the addition of Mg2+ to the growth medium. RMF deficiency was found to inhibit the synthesis of another cation-selective porin OmpF. Thus, the double rmf ompC mutant is deficient in both OmpC and OmpF, which probably accounts for the pronounced decrease in Mg2+ uptake in this mutant. The results indicate that both RMF and Mg2+, acting through stabilization of ribosomes, are important for cell viability at the stationary growth phase.
核糖体调控因子(RMF)在大肠杆菌从指数生长期向稳定生长期转变的过程中参与核糖体的稳定。已知RMF缺乏会降低细胞活力。亚精胺的过度积累也会导致细胞活力下降以及RMF和阳离子选择性孔蛋白OmpC的合成减少。因此,RMF水平的降低可能与亚精胺过量导致的细胞活力下降有关。由于亚精胺也会影响OmpC的表达,我们研究了OmpC缺乏是否会增强由RMF缺乏引起的细胞死亡。ompC突变体本身并不影响蛋白质合成或细胞活力,但rmf ompC双突变体导致的蛋白质合成和细胞活力下降比单一的rmf突变体大得多。rmf ompC双突变体中的核糖体数量和Mg2+含量也有所下降,向生长培养基中添加Mg2+可部分恢复细胞活力。发现RMF缺乏会抑制另一种阳离子选择性孔蛋白OmpF的合成。因此,rmf ompC双突变体同时缺乏OmpC和OmpF,这可能是该突变体中Mg2+摄取显著减少的原因。结果表明,RMF和Mg2+通过稳定核糖体发挥作用,对稳定生长期的细胞活力都很重要。