title: General and condition-specific essential functions of Pseudomonas aeruginosa.
abstract: The essential functions of a bacterial pathogen reflect the most basic processes required for its viability and growth, and represent potential therapeutic targets. Most screens for essential genes have assayed a single condition--growth in a rich undefined medium--and thus have not distinguished genes that are generally essential from those that are specific to this particular condition. To help define these classes for Pseudomonas aeruginosa, we identified genes required for growth on six different media, including a medium made from cystic fibrosis patient sputum. The analysis used the Tn-seq circle method to achieve high genome coverage and analyzed more than 1,000,000 unique insertion positions (an average of one insertion every 6.0 bp). We identified 352 general and 199 condition-specific essential genes. A subset of assignments was verified in individual strains with regulated expression alleles. The profile of essential genes revealed that, compared with Escherichia coli, P. aeruginosa is highly vulnerable to mutations disrupting central carbon-energy metabolism and reactive oxygen defenses. These vulnerabilities may arise from the stripped-down architecture of the organism's carbohydrate utilization pathways and its reliance on respiration for energy generation. The essential function profile thus provides fundamental insights into P. aeruginosa physiology as well as identifying candidate targets for new antibacterial agents.
authors.affiliation: Department of Genome Sciences and.
authors.forename: Samuel A
authors.initials: SA
authors.lastname: Lee
authors.affiliation: Department of Genome Sciences and.
authors.forename: Larry A
authors.initials: LA
authors.lastname: Gallagher
authors.affiliation: Center for Zoonotic Diseases, Faculty of Veterinary Science, Mahidol University, 73170 Bangkok, Thailand.
authors.forename: Metawee
authors.initials: M
authors.lastname: Thongdee
authors.affiliation: Department of Microbiology and Medicine, University of Washington, Seattle, WA 98195; and.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Staudinger
authors.affiliation: Department of Genome Sciences and.
authors.forename: Soyeon
authors.initials: S
authors.lastname: Lippman
authors.affiliation: Department of Microbiology and Medicine, University of Washington, Seattle, WA 98195; and.
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Singh
authors.affiliation: Department of Genome Sciences and manoil@u.washington.edu.
authors.forename: Colin
authors.initials: C
authors.lastname: Manoil
mesh_terms: D002244:Carbon / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D020043:Genes, Essential; D016254:Mutagenesis, Insertional / Q000235:genetics; D010802:Phylogeny; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D002244:Carbon
keywords: ESKAPE; Tn-seq; antibiotic target; cystic fibrosis; sputum
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