To successfully infect humans, Pseudomonas aeruginosa (Pa) must overcome the low iron availability in host tissues. A transcriptome comparison was carried out between iron-starved cells of Pa treated with iron and untreated controls. The present study is the first global analysis of the early transcriptional response of exponentially growing Pa to iron. Approximately 1.3% of the Pa genes displayed > or = 5.0-fold changes in mRNA levels in iron-treated cells. Treatment affected the mRNA levels of many genes required for iron acquisition as well as several genes with relevance to virulence previously known to be regulated by iron. More importantly, the analysis permitted identification of 107 Pa genes whose mRNA levels were not previously known to be affected by iron. These genes are good candidates for mutagenesis studies aimed at identifying novel functions relevant to iron metabolism in Pa. Some of these genes encode predicted siderophore receptors, iron transport systems, TonB-dependent receptors, regulatory proteins, and proteins relevant to virulence. Notably, 49 genes encode hypothetical or conserved hypothetical proteins of unknown function, suggesting that they are involved directly or indirectly in iron metabolism or metabolic adaptation to different iron-availability conditions.