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Proc Natl Acad Sci U S A. 2015 Mar 31;112(13):4110-5. doi: 10.1073/pnas.1419677112. Epub 2015 Mar 16.

Essential genome of Pseudomonas aeruginosa in cystic fibrosis sputum.

Author information

1
Department of Molecular Biosciences, Institute of Cellular and Molecular Biology, Center for Infectious Disease, The University of Texas at Austin, Austin, TX 78712.
2
Department of Molecular Biosciences, Institute of Cellular and Molecular Biology, Center for Infectious Disease, The University of Texas at Austin, Austin, TX 78712 mwhiteley@austin.utexas.edu.

Abstract

Defining the essential genome of bacterial pathogens is central to developing an understanding of the biological processes controlling disease. This has proven elusive for Pseudomonas aeruginosa during chronic infection of the cystic fibrosis (CF) lung. In this paper, using a Monte Carlo simulation-based method to analyze high-throughput transposon sequencing data, we establish the P. aeruginosa essential genome with statistical precision in laboratory media and CF sputum. Reconstruction of the global requirements for growth in CF sputum compared with defined growth conditions shows that the latter requires several cofactors including biotin, riboflavin, and pantothenate. Comparison of P. aeruginosa strains PAO1 and PA14 demonstrates that essential genes are primarily restricted to the core genome; however, some orthologous genes in these strains exhibit differential essentiality. These results indicate that genes with similar molecular functions may have distinct genetic roles in different P. aeruginosa strains during growth in CF sputum. We also show that growth in a defined growth medium developed to mimic CF sputum yielded virtually identical fitness requirements to CF sputum, providing support for this medium as a relevant in vitro model for CF microbiology studies.

KEYWORDS:

Pseudomonas aeruginosa; Tn-seq; cystic fibrosis; essential genes; sputum

PMID:
25775563
PMCID:
PMC4386324
DOI:
10.1073/pnas.1419677112
[Indexed for MEDLINE]
Free PMC Article

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