In vitro and in vivo screening for novel essential cell-envelope proteins in Pseudomonas aeruginosa

Regina Fernandez-Pinar, Alessandra Lo Sciuto, Alice Rossi, Serena Ranucci, Alessandra Bragonzi, Francesco Imperi

Research output: Contribution to journalArticlepeer-review


The Gram-negative bacterium Pseudomonas aeruginosa represents a prototype of multi-drug resistant opportunistic pathogens for which novel therapeutic options are urgently required. In order to identify new candidates as potential drug targets, we combined large-scale transposon mutagenesis data analysis and bioinformatics predictions to retrieve a set of putative essential genes which are conserved in P. aeruginosa and predicted to encode cell envelope or secreted proteins. By generating unmarked deletion or conditional mutants, we confirmed the in vitro essentiality of two periplasmic proteins, LptH and LolA, responsible for lipopolysaccharide and lipoproteins transport to the outer membrane respectively, and confirmed that they are important for cell envelope stability. LptH was also found to be essential for P. aeruginosa ability to cause infection in different animal models. Conversely, LolA-depleted cells appeared only partially impaired in pathogenicity, indicating that this protein likely plays a less relevant role during bacterial infection. Finally, we ruled out any involvement of the other six proteins under investigation in P. aeruginosa growth, cell envelope stability and virulence. Besides proposing LptH as a very promising drug target in P. aeruginosa, this study confirms the importance of in vitro and in vivo validation of potential essential genes identified through random transposon mutagenesis.

Original languageEnglish
Article number17593
JournalScientific Reports
Publication statusPublished - Dec 1 2015

ASJC Scopus subject areas

  • General


Dive into the research topics of 'In vitro and in vivo screening for novel essential cell-envelope proteins in Pseudomonas aeruginosa'. Together they form a unique fingerprint.

Cite this