Pang H, Winkler H H
Department of Microbiology and Immunology, University of South Alabama College of Medicine, Mobile 36688.
Mol Microbiol. 1994 Apr;12(1):115-20. doi: 10.1111/j.1365-2958.1994.tb01000.x.
The obligate intracellular parasite, Rickettsia prowazekii, is a slow-growing bacterium with a doubling time of about 10 h. In the present study, DNA and RNA were obtained from the rickettsiae by two independent methods, i.e. simultaneous isolation of DNA and RNA from the same sample by phenol:chloroform extraction and CsCI gradient centrifugation. In addition, ribosomal RNA was obtained by sedimentation of partially purified ribosomes from the rickettsiae. The results demonstrated that, after correction for the cell volumes, the concentrations of stable RNA and ribosomes in R. prowazekii, a slow-growing organism, were about 62 fg micron-3 and 17,000 per micron3, respectively, which were very similar (66 fg micron-3 and 21,000 per micron3) to those in Escherichia coli with a generation time of 40 min. However, on a per cell basis, R. prowazekii had 5.6 fg of RNA and 1500 ribosomes per cell, which was only about 8% of the amount of both stable RNA (71.2 fg) and ribosomes (24,000) per cell as was found in E. coli. These results indicated that R. prowazekii possesses a ribosome concentration greater than might have been predicted from its slow growth rate. This high concentration of ribosomes could be due to a large population of nonfunctioning ribosomes, a low efficiency of amino acid production, or a high rate of protein turnover. However, this study also demonstrated that the rickettsiae have very limited protein turnover. Knowledge of the kinetics and control mechanisms for protein synthesis in R. prowazekii remains to be established to determine the logic of the extra rickettsial ribosomes.
专性胞内寄生虫普氏立克次氏体是一种生长缓慢的细菌,其倍增时间约为10小时。在本研究中,通过两种独立方法从立克次氏体中获得DNA和RNA,即通过苯酚:氯仿提取和氯化铯梯度离心从同一样品中同时分离DNA和RNA。此外,通过沉降立克次氏体中部分纯化的核糖体获得核糖体RNA。结果表明,校正细胞体积后,生长缓慢的普氏立克次氏体中稳定RNA和核糖体的浓度分别约为62 fg/μm³和17,000个/μm³,这与世代时间为40分钟的大肠杆菌中的浓度(66 fg/μm³和21,000个/μm³)非常相似。然而,以每个细胞计算,普氏立克次氏体每个细胞有5.6 fg的RNA和1500个核糖体,这仅约为大肠杆菌中每个细胞稳定RNA(71.2 fg)和核糖体(24,000个)数量的8%。这些结果表明,普氏立克次氏体的核糖体浓度高于根据其缓慢生长速率可能预测的浓度。核糖体的这种高浓度可能是由于大量无功能的核糖体、氨基酸产生效率低或蛋白质周转率高。然而,本研究还表明立克次氏体的蛋白质周转率非常有限。普氏立克次氏体中蛋白质合成的动力学和控制机制的知识仍有待确定,以弄清楚立克次氏体额外核糖体的逻辑。