title: Direct observation of single stationary-phase bacteria reveals a surprisingly long period of constant protein production activity.
abstract: Exponentially growing bacteria are rarely found in the wild, as microorganisms tend to spend most of their lifetime at stationary phase. Despite this general prevalence of stationary-phase bacteria, they are as yet poorly characterized. Our goal was to quantitatively study this phase by direct observation of single bacteria as they enter into stationary phase and by monitoring their activity over several days during growth arrest. For this purpose, we devised an experimental procedure for starving single Escherichia coli bacteria in microfluidic devices and measured their activity by monitoring the production rate of fluorescent proteins. When amino acids were the sole carbon source, the production rate decreased by an order of magnitude upon entry into stationary phase. We found that, even while growth-arrested, bacteria continued to produce proteins at a surprisingly constant rate over several days. Our identification of this newly observed period of constant activity in nongrowing cells, designated as constant activity stationary phase, makes possible the conduction of assays that require constant protein expression over time, and are therefore difficult to perform under exponential growth conditions. Moreover, we show that exogenous protein expression bears no fitness cost on the regrowth of the population when starvation ends. Further characterization of constant activity stationary phase-a phase where nongrowing bacteria can be quantitatively studied over several days in a reproducible manner-should contribute to a better understanding of this ubiquitous but overlooked physiological state of bacteria in nature.
authors.affiliation: Racah Institute for Physics, and Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem, Jerusalem 9270001, Israel.
authors.forename: Orit
authors.initials: O
authors.lastname: Gefen
authors.forename: Ofer
authors.initials: O
authors.lastname: Fridman
authors.forename: Irine
authors.initials: I
authors.lastname: Ronin
authors.forename: Nathalie Q
authors.initials: NQ
authors.lastname: Balaban
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial; D046210:Microfluidic Analytical Techniques; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins
keywords: CASP; bacterial metabolism; constant activity stationary phase