Bacteria growing in different conditions experience a broad range of demand on the rate of protein synthesis which profoundly affects cellular resource allocation. During fast growth, protein synthesis is long known to be modulated by adjusting the ribosome content, with the vast majority of ribosomes engaged at a near-maximal rate of elongation. Here we characterized protein synthesis by E. coli systematically, focusing on slow growth conditions. We establish that the translational elongation rate decreases as growth slows down, exhibiting a Michaelis-Menten dependence on the abundance of the cellular translational apparatus. However, an appreciable elongation rate is maintained even towards zero growth including the stationary phase. This maintenance, critical for timely protein synthesis in harsh environments, is accompanied by a drastic reduction in the fraction of active ribosomes. Interestingly, well-known antibiotics such as chloramphenicol also cause substantial reduction in the pool of active ribosomes, instead of slowing down translational elongation as commonly thought.

Understanding how the homeostasis of cellular size and composition is accomplished by different organisms is an outstanding challenge in biology. For exponentially growing Escherichia coli cells, it is long known that the size of cells exhibits a strong positive relation with their growth rates in different nutrient conditions. Here, we characterized cell sizes in a set of orthogonal growth limitations. We report that cell size and mass exhibit positive or negative dependences with growth rate depending on the growth limitation applied. In particular, synthesizing large amounts of “useless” proteins led to an inversion of the canonical, positive relation, with slow growing cells enlarged 7- to 8-fold compared to cells growing at similar rates under nutrient limitation. Strikingly, this increase in cell size was accompanied by a 3- to 4-fold increase in cellular DNA content at slow growth, reaching up to an amount equivalent to ∼8 chromosomes per cell. Despite drastic changes in cell mass and macromolecular composition, cellular dry mass density remained constant. Our findings reveal an important role of protein synthesis in cell division control.

Introductory paragraph Tight coordination between transcription and translation is crucial to maintaining the integrity of gene expression in bacteria. How bacteria manage to coordinate these two processes remains unclear. Possible direct physical coupling between the RNA polymerase and ribosome has been intensely investigated in recent years. Here, we quantitatively characterize the transcriptional kinetics of E. coli in different growth conditions. Transcriptional and translational elongation remain coordinated under various nutrient conditions as previously reported. However, transcriptional elongation was not affected under antibiotics that slowed down translational elongation. The same was found by introducing nonsense mutation that completely dissociated transcription from translation. Our data thus provide direct evidences showing that translation is not required to maintain the speed of transcriptional elongation. In cases where transcription and translation are dissociated, our study provides quantitative characterization of the resulting process of premature transcriptional termination (PTT). PTT-mediated polarity caused by translationtargeting antibiotics substantially affected the coordinated expression of genes in several long operons studied, contributing to important physiological effects of these antibiotics. Our results also suggest a model in which the coordination between transcriptional and translational elongation under normal growth conditions is implemented by ppGpp.