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Identification of Fitness Determinants during Energy-Limited Growth Arrest in Pseudomonas aeruginosa

Basta, David W. and Bergkessel, Megan and Newman, Dianne K. (2017) Identification of Fitness Determinants during Energy-Limited Growth Arrest in Pseudomonas aeruginosa. mBio, 8 (6). Art. No. e01170. ISSN 2150-7511. PMCID PMC5705914; PMC5893874.

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Microbial growth arrest can be triggered by diverse factors, one of which is energy limitation due to scarcity of electron donors or acceptors. Genes that govern fitness during energy-limited growth arrest and the extent to which they overlap between different types of energy limitation are poorly defined. In this study, we exploited the fact that Pseudomonas aeruginosa can remain viable over several weeks when limited for organic carbon (pyruvate) as an electron donor or oxygen as an electron acceptor. ATP values were reduced under both types of limitation, yet more severely in the absence of oxygen. Using transposon-insertion sequencing (Tn-seq), we identified fitness determinants in these two energy-limited states. Multiple genes encoding general functions like transcriptional regulation and energy generation were required for fitness during carbon or oxygen limitation, yet many specific genes, and thus specific activities, differed in their relevance between these states. For instance, the global regulator RpoS was required during both types of energy limitation, while other global regulators such as DksA and LasR were required only during carbon or oxygen limitation, respectively. Similarly, certain ribosomal and tRNA modifications were specifically required during oxygen limitation. We validated fitness defects during energy limitation using independently generated mutants of genes detected in our screen. Mutants in distinct functional categories exhibited different fitness dynamics: regulatory genes generally manifested a phenotype early, whereas genes involved in cell wall metabolism were required later. Together, these results provide a new window into how P. aeruginosa survives growth arrest.

Item Type:Article
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URLURL TypeDescription CentralArticle CentralErratum
Basta, David W.0000-0003-4176-6566
Bergkessel, Megan0000-0002-4530-1224
Newman, Dianne K.0000-0003-1647-1918
Other Contributors:
ContributionOther Contributors NameIdentifierPersonID (may be blank)
EditorWhiteley, MarvinWhiteley-MUNSPECIFIED
Additional Information:© 2017 Basta et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Received 31 July 2017. Accepted 13 October 2017. Published 28 November 2017. We thank Larry Gallagher for advice on generating the transposon library, preparing samples for Tn-seq, and the gift of plasmid pIT2, Heather Curtis for the illustration in Fig. 2A, Igor Antoshechkin for optimization and troubleshooting of sequencing, Gargi Kulkarni for advice and troubleshooting of cloning, Nate Glasser for assistance with HPLC, and members of the Newman lab for support and constructive feedback on the manuscript. This study was supported by NIH grant 5R01HL117328-03 to D.K.N.
Errata:Volume 8, no. 6, e01170-17, 2017, The following should be added to the end of Acknowledgments: “This study was supported by NIH grants 5R01AI127850 and 5R01HL117328 to D.K.N.”
Funding AgencyGrant Number
Subject Keywords:Pseudomonas aeruginosa, Tn-seq, fitness, growth arrest, slow growth, energy limitation
Issue or Number:6
PubMed Central ID:PMC5705914; PMC5893874
Record Number:CaltechAUTHORS:20171205-204626357
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83718
Deposited By: Donna Wrublewski
Deposited On:06 Dec 2017 23:17
Last Modified:14 Apr 2020 20:40

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