Acinetobacter baumannii is a common pathogen whose recent resistance to drugs has emerged as a major health problem. Ethanol has been found to increase the virulence of A. baumannii in Dictyostelium discoideum and Caenorhabditis elegans models of infection. To better understand the causes of this effect, we examined the transcriptional profile of A. baumannii grown in the presence or absence of ethanol using RNA-Seq. Using the Illumina/Solexa platform, a total of 43,453,960 reads (35 nt) were obtained, of which 3,596,474 mapped uniquely to the genome. Our analysis revealed that ethanol induces the expression of 49 genes that belong to different functional categories. A strong induction was observed for genes encoding metabolic enzymes, indicating that ethanol is efficiently assimilated. In addition, we detected the induction of genes encoding stress proteins, including upsA, hsp90, groEL and lon as well as permeases, efflux pumps and a secreted phospholipase C. In stationary phase, ethanol strongly induced several genes involved with iron assimilation and a high-affinity phosphate transport system, indicating that A. baumannii makes a better use of the iron and phosphate resources in the medium when ethanol is used as a carbon source. To evaluate the role of phospholipase C (Plc1) in virulence, we generated and analyzed a deletion mutant for plc1. This strain exhibits a modest, but reproducible, reduction in the cytotoxic effect caused by A. baumannii on epithelial cells, suggesting that phospholipase C is important for virulence. Overall, our results indicate the power of applying RNA-Seq to identify key modulators of bacterial pathogenesis. We suggest that the effect of ethanol on the virulence of A. baumannii is multifactorial and includes a general stress response and other specific components such as phospholipase C.
Acinetobacter baumannii has recently emerged as a frequent opportunistic pathogen. In the presence of ethanol A. baumannii increases its pathogenicity towards Dictyostelium discoideum and Caenorhabditis elegans, and community-acquired infections of A. baumannii are associated with alcoholism. Ethanol negatively affects both epithelial cells and alters the bacterial physiology. To explore the underlying basis for the increased virulence of A. baumannii in the presence of ethanol we examined the transcriptional profile of this bacterium using the novel methodology known as RNA-Seq. We show that ethanol induces the expression of a phospholipase C, which contributes to A. baumannii cytotoxicity. We also show that many proteins related to stress were induced and that ethanol is efficiently assimilated as a carbon source leading to induction in stationary phase of two different Fe uptake systems and a phosphate transport system. Interestingly, a previous study showed that a mutant in the high-affinity phosphate uptake system was avirulent. Our work contributes to the understanding of A. baumannii pathogenesis and provides a powerful approach that can be extended to other pathogenic bacteria.