Molecular and Computational Biology Section, Department of Biological Sciences, University of Southern Californiagrid.42505.36, Los Angeles, California, USA.
Microbiol Spectr. 2022 Apr 27;10(2):e0223921. doi: 10.1128/spectrum.02239-21. Epub 2022 Apr 4.
During stationary phase in Escherichia coli, the expression of the ribosome modulation factor (RMF) protein participates in the dimerization of two 70S ribosomes, ultimately creating a 100S particle. 100S ribosomes are commonly thought to function to preserve ribosomes as growth ceases and cells begin to catabolize intracellular components, including proteins, during their transition into stationary phase. Here, we show that the rates of stationary-phase ribosomal degradation are increased in an mutant strain that cannot produce 100S ribosomes, resulting in deficiencies in outgrowth upon reinoculation into fresh medium. Upon coinoculation in LB medium, the mutant exhibits a delay in entry into log phase, differences in growth rates, and an overall reduction in relative fitness during competition. Unexpectedly, the mutant exhibited shorter generation times than wild-type cells during log phase, both in monoculture and during competition. These doubling times of ∼13 min suggest that failure to maintain ribosomal balance affects the control of cell division. Though the timing of entry into and exit from log phase is altered, 100S ribosomes are not essential for long-term viability of the mutant when grown in monoculture. Ribosomes are the sole source in any cell for new protein synthesis that is vital to maintain life. While ribosomes are frequently consumed as sources of nutrients under low-nutrient conditions, some ribosomes appear to be preserved for later use. The failure to maintain the availability of these ribosomes can lead to a dire consequence upon the influx of new nutrients, as cells are unable to efficiently replenish their metabolic machinery. It is important to study the repercussions, consequences, and mechanisms of survival in cells that cannot properly maintain the availability of their ribosomes in order to better understand their mechanisms of survival during competition under nutrient-depleted conditions.
在大肠杆菌的静止期,核糖体调节因子(RMF)蛋白的表达参与两个 70S 核糖体的二聚化,最终形成 100S 颗粒。100S 核糖体通常被认为是在生长停止时起作用的,当细胞开始分解细胞内的成分(包括蛋白质)时,这些核糖体作为细胞进入静止期的一种保护机制。在这里,我们表明,在不能产生 100S 核糖体的 突变菌株中,静止期核糖体降解的速度增加,导致在重新接种到新鲜培养基中时,出芽能力不足。在 LB 培养基中共同接种时,该突变体在进入对数期时表现出延迟,在生长速度上存在差异,并且在竞争中相对适应度总体降低。出乎意料的是,与野生型细胞相比,在对数期,突变体在单培养和竞争中表现出较短的代时。这些倍增时间约为 13 分钟表明,核糖体平衡的失败会影响细胞分裂的控制。尽管进入和退出对数期的时间发生了变化,但在单培养中,当生长时,100S 核糖体对于 突变体的长期存活并不是必需的。核糖体是任何细胞中新蛋白质合成的唯一来源,这对于维持生命至关重要。虽然核糖体在低营养条件下经常被用作营养物质的来源,但一些核糖体似乎被保留下来以备后用。如果不能维持这些核糖体的可用性,当新的营养物质涌入时,细胞将无法有效地补充其代谢机制,这将导致严重的后果。研究不能正确维持核糖体可用性的细胞的后果、影响和生存机制,对于更好地理解它们在营养物质耗尽条件下竞争时的生存机制非常重要。