ABSTRACT Heterotrophic bacteria rapidly deplete essential macronutrients during growth and must navigate subsequent periods of growth arrest imposed by starvation. Nutrient limitations can be dynamic in nature, requiring ongoing regulatory adjustments involving new protein synthesis despite total biosynthetic activities being dramatically lower than during growth. Here, we have characterized the responses of the opportunistic pathogen Pseudomonas aeruginosa to prolonged starvation for carbon or nitrogen sources and to transitions between these states. We find that most cells survive both types of starvation for more than a week and maintain low but robustly detectable levels of protein synthesis in the absence of growth. Nitrogen-starved cells are larger, make more proteins, and retain fewer ribosomes than carbon-starved cells, indicating that distinct physiological strategies are adopted during the two starvation types. We found that the newly synthesized proteomes of each starvation type are distinct although many of the most highly synthesized proteins are shared between both conditions. Interestingly, we observed a temporary burst of protein synthesis as cells were transitioned between the two starvation conditions, which may reflect active remodeling of the proteome during growth arrest. We also used transposon insertion sequencing to identify genes impacting fitness in both starvation conditions and during transitions between the two and found that a highly overlapping set of global regulators most strongly influenced survival. Combining these data sets, we highlight proteases and chaperones, flagellar motility, and the nitrogen-related phosphotransferase system as key fitness-impacting functions that are actively maintained by growth-arrested Pseudomonas aeruginosa . IMPORTANCE Molecular microbiology has traditionally focused on exponential growth in model organisms as the preferred context in which to study bacterial physiology, especially the regulation of new protein synthesis. However, in natural environments, including many infection contexts, bacteria frequently enter growth arrest due to nutrient limitation. The dynamics and regulation of protein synthesis in growth-arrested cells remain poorly understood, especially in pathogens. Furthermore, growth arrest increases tolerance to a variety of stresses, including many clinically used antimicrobials. We have conducted a comprehensive exploration of the proteins being made by growth arrested Pseudomonas aeruginosa during total nitrogen or carbon starvation and at the transition between these two starvation types, and the genes supporting fitness under these conditions. These datasets suggest dynamic redistribution of resources among important cellular functions and will serve as a resource for further investigations of starvation-induced growth arrest, a ubiquitous but understudied physiological state of heterotrophic bacteria.
Munro et al. (Fri,) studied this question.