General and condition-specific essential functions ofPseudomonas 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 forPseudomonas 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 withEscherichia coli,P. aeruginosais 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 intoP. aeruginosaphysiology as well as identifying candidate targets for new antibacterial agents.