Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Single cell fitness of B. subtilis correlates with the ability to adapt to various stresses.

ABSTRACT: How cells dynamically respond to fluctuating environmental conditions depends on the architecture and noise of the underlying genetic circuits. Most work characterizing stress pathways in the model bacterium Bacillus subtilis has been performed on bulk cultures using ensemble assays. However, investigating the single cell response to stress is important since noise might generate significant phenotypic heterogeneity. Here, we study a single stress response (glucose starvation) to allow comparison of both population- and single cell data. Using a top-down approach, we investigate the transcriptional dynamics of various stress-related genes of B. subtilis in response to glucose starvation. Our data reveal that most stress-induced pathways respond highly heterogeneously to glucose starvation. Bacteria are commonly thought to exhibit phenotypic heterogeneity to increase their survival under changing environments by always having a subpopulation pre-adapted to adverse conditions. Interestingly, growth experiments with cultures that were either pre-adapted for glucose starvation or non-starved cells showed that non-starved cells demonstrate a significant growth advantage when challenged with a new stress. This suggests that once cells engage in a certain stress pathway they are only optimised for this specific condition. Hence it is important for the population to always maintain a 'naM-CM-/ve' subpopulation in case of fluctuating environments. Our data indicate the presence of performance-based selection, by which only cells with a sufficiently high-energy status or cellular fitness are able to diversify. The transcriptome of glucose-starved and non-starved B. subtilis 168 trp+ cultures was compared using 3 biological replicates.

ORGANISM(S): Bacillus subtilis

SUBMITTER: Imke de Jong

PROVIDER: E-GEOD-33506 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Extended periods of waking result in physiological impairments in humans, rats, and flies. Sleep homeostasis, the increase in sleep observed following sleep loss, is believed to counter the negative effects of prolonged waking by restoring vital biological processes that are degraded during sleep deprivation. Sleep homeostasis, as with other behaviors, is influenced by both genes and environment. We report here that during periods of starvation, flies remain spontaneously awake but, in contrast to sleep deprivation, do not accrue any of the negative consequences of prolonged waking. Specifically, the homeostatic response and learning impairments that are a characteristic of sleep loss are not observed following prolonged waking induced by starvation. To identify the genes responsible for the protective effects of starvation we conducted transcription profiling of sleep deprived flies that accrue sleep debt compared to starved siblings that do not. Genes involved in lipid metabolism were highly enriched in our dataset of 84 differentially regulated transcripts. Follow up genetic studies established that 6 genes involved in lipid metabolism strongly influence sleep homeostasis. Two of these genes, brummer (bmm) and Lipid storage droplet 2 (Lsd2), are in the same lipolysis pathway but exert antagonistic effects on lipid storage. bmm mutant flies have excess fat stores and display a large homeostatic response following sleep deprivation. In contrast, Lsd2 mutant flies, which phenocopy aspects of starvation as measured by low triglyceride stores, do not exhibit a homeostatic response following sleep loss. Importantly, Lsd2 mutant flies are not learning impaired after sleep deprivation. These results provide the first genetic evidence, to our knowledge, that lipid metabolism plays an important role in regulating the homeostatic response and can protect against neuronal impairments induced by prolonged waking. Two-condition experiments: sleep deprived vs starved. RNA from 8 biological replicates for each condition was pooled in groups of 2 to create 4 samples. Each of the 4 samples is run in duplicate with untreated circadian matched controls.

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Single cell fitness of B. subtilis correlates with the ability to adapt to various stresses.

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