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Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth

Population-level analyses are rapidly becoming inadequate to answer many of biomedical science and microbial ecology’s most pressing questions. The role of microbial populations within ecosystems and the evolutionary selective pressure on individuals depend fundamentally on the metabolic activity of...

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Autores principales: McClelland, H. L. O., Jones, C., Chubiz, L. M., Fike, D. A., Bradley, A. S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7064762/
https://www.ncbi.nlm.nih.gov/pubmed/32127448
http://dx.doi.org/10.1128/mBio.01519-19
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author McClelland, H. L. O.
Jones, C.
Chubiz, L. M.
Fike, D. A.
Bradley, A. S.
author_facet McClelland, H. L. O.
Jones, C.
Chubiz, L. M.
Fike, D. A.
Bradley, A. S.
author_sort McClelland, H. L. O.
collection PubMed
description Population-level analyses are rapidly becoming inadequate to answer many of biomedical science and microbial ecology’s most pressing questions. The role of microbial populations within ecosystems and the evolutionary selective pressure on individuals depend fundamentally on the metabolic activity of single cells. Yet, many existing single-cell technologies provide only indirect evidence of metabolic specialization because they rely on correlations between transcription and phenotype established at the level of the population to infer activity. In this study, we take a top-down approach using isotope labels and secondary ion mass spectrometry to track the uptake of carbon and nitrogen atoms from different sources into biomass and directly observe dynamic changes in anabolic specialization at the level of single cells. We investigate the classic microbiological phenomenon of diauxic growth at the single-cell level in the model methylotroph Methylobacterium extorquens. In nature, this organism inhabits the phyllosphere, where it experiences diurnal changes in the available carbon substrates, necessitating an overhaul of central carbon metabolism. We show that the population exhibits a unimodal response to the changing availability of viable substrates, a conclusion that supports the canonical model but has thus far been supported by only indirect evidence. We anticipate that the ability to monitor the dynamics of anabolism in individual cells directly will have important applications across the fields of ecology, medicine, and biogeochemistry, especially where regulation downstream of transcription has the potential to manifest as heterogeneity that would be undetectable with other existing single-cell approaches.
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spelling pubmed-70647622020-03-13 Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth McClelland, H. L. O. Jones, C. Chubiz, L. M. Fike, D. A. Bradley, A. S. mBio Observation Population-level analyses are rapidly becoming inadequate to answer many of biomedical science and microbial ecology’s most pressing questions. The role of microbial populations within ecosystems and the evolutionary selective pressure on individuals depend fundamentally on the metabolic activity of single cells. Yet, many existing single-cell technologies provide only indirect evidence of metabolic specialization because they rely on correlations between transcription and phenotype established at the level of the population to infer activity. In this study, we take a top-down approach using isotope labels and secondary ion mass spectrometry to track the uptake of carbon and nitrogen atoms from different sources into biomass and directly observe dynamic changes in anabolic specialization at the level of single cells. We investigate the classic microbiological phenomenon of diauxic growth at the single-cell level in the model methylotroph Methylobacterium extorquens. In nature, this organism inhabits the phyllosphere, where it experiences diurnal changes in the available carbon substrates, necessitating an overhaul of central carbon metabolism. We show that the population exhibits a unimodal response to the changing availability of viable substrates, a conclusion that supports the canonical model but has thus far been supported by only indirect evidence. We anticipate that the ability to monitor the dynamics of anabolism in individual cells directly will have important applications across the fields of ecology, medicine, and biogeochemistry, especially where regulation downstream of transcription has the potential to manifest as heterogeneity that would be undetectable with other existing single-cell approaches. American Society for Microbiology 2020-03-03 /pmc/articles/PMC7064762/ /pubmed/32127448 http://dx.doi.org/10.1128/mBio.01519-19 Text en Copyright © 2020 McClelland et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Observation
McClelland, H. L. O.
Jones, C.
Chubiz, L. M.
Fike, D. A.
Bradley, A. S.
Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth
title Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth
title_full Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth
title_fullStr Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth
title_full_unstemmed Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth
title_short Direct Observation of the Dynamics of Single-Cell Metabolic Activity during Microbial Diauxic Growth
title_sort direct observation of the dynamics of single-cell metabolic activity during microbial diauxic growth
topic Observation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7064762/
https://www.ncbi.nlm.nih.gov/pubmed/32127448
http://dx.doi.org/10.1128/mBio.01519-19
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