Makinoshima Hideki, Aizawa Shin-Ichi, Hayashi Hideo, Miki Takeyoshi, Nishimura Akiko, Ishihama Akira
Division of Molecular Biology, Nippon Institute for Biological Science, Ome, Tokyo 198-0024, Japan.
J Bacteriol. 2003 Feb;185(4):1338-45. doi: 10.1128/JB.185.4.1338-1345.2003.
Escherichia coli cultures can be fractionated into more than 20 cell populations, each having a different bouyant density and apparently representing a specific stage of cell differentiation from exponential growth to stationary phase (H. Makinoshima, A. Nishimura, and A. Ishihama, Mol. Microbiol. 43:269-279, 2002). The density increase was found to be impaired at an early step for a mutant E. coli with the disrupted rpoS gene, which encodes the RNA polymerase RpoS (sigma-S) for stationary-phase gene transcription. This finding suggests that RpoS is need for the entire process of cell density increase. In the absence of RpoF sigma factor, the flagella are not formed as observed by electron microscopy, but the growth phase-coupled density increase takes place as in wild-type E. coli, confirming that the alteration in cell density is not directly correlated with the presence or absence of flagella. In the stationary-phase cells, accumulation of electron-dense areas was observed by electron microscopic observation of bacterial thin sections. By chemical determination, the increase in glycogen (or polysaccharides) was suggested to be one component, which contributes to the increase in weight-to-volume ratio of stationary-phase E. coli cells.
大肠杆菌培养物可被分离成20多个细胞群体,每个群体具有不同的浮力密度,显然代表了从指数生长期到稳定期细胞分化的特定阶段(H. Makinoshima、A. Nishimura和A. Ishihama,《分子微生物学》43:269 - 279,2002年)。对于编码用于稳定期基因转录的RNA聚合酶RpoS(σ - S)的rpoS基因被破坏的突变型大肠杆菌,发现其在早期阶段密度增加受到损害。这一发现表明RpoS是细胞密度增加整个过程所必需的。在没有RpoF σ因子的情况下,通过电子显微镜观察发现鞭毛未形成,但生长阶段耦合的密度增加与野生型大肠杆菌一样发生,这证实了细胞密度的改变与鞭毛的有无没有直接关联。在稳定期细胞中,通过对细菌薄片的电子显微镜观察,观察到了电子致密区域的积累。通过化学测定,有人提出糖原(或多糖)的增加是一个因素,它有助于稳定期大肠杆菌细胞重量与体积比的增加。