The tyrosine kinase BceF and the phosphotyrosine phosphatase BceD of Burkholderia contaminans are required for efficient invasion and epithelial disruption of a cystic fibrosis lung epithelial cell line

• Published in: Infection and immunity 2015-02, Vol.83 (2), p.812-821
• Main Authors: Ferreira, Ana S, Silva, Inês N, Fernandes, Fábio, Pilkington, Ruth, Callaghan, Máire, McClean, Siobhán, Moreira, Leonilde M
• Format: Article
• Language: English
• Subjects: Albumins - metabolism; Anti-Bacterial Agents - pharmacology; Bacterial Adhesion; Burkholderia; Burkholderia cepacia complex - enzymology; Burkholderia cepacia complex - genetics; Burkholderia cepacia complex - pathogenicity; Burkholderia Infections - microbiology; Burkholderia Infections - pathology; Cell Line; Cellular Microbiology: Pathogen-Host Cell Molecular Interactions; Ciprofloxacin - pharmacology; Claudin-1 - biosynthesis; Cystic Fibrosis - microbiology; Cystic Fibrosis Transmembrane Conductance Regulator - genetics; Electric Impedance; Humans; Inflammation - immunology; Interleukin-6 - biosynthesis; Interleukin-6 - metabolism; Interleukin-8 - biosynthesis; Interleukin-8 - metabolism; Lung - microbiology; Membrane Potentials; Mutation; Occludin - biosynthesis; Protein Transport; Protein Tyrosine Phosphatases - genetics; Protein Tyrosine Phosphatases - physiology; Protein-Tyrosine Kinases - genetics; Protein-Tyrosine Kinases - physiology; Respiratory Mucosa - microbiology; Tight Junctions - pathology; Virulence Factors - genetics; Zonula Occludens-1 Protein - biosynthesis
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/IAI.02713-14

Bacterial tyrosine kinases and their cognate protein tyrosine phosphatases are best known for regulating the biosynthesis of polysaccharides. Moreover, their roles in the stress response, DNA metabolism, cell division, and virulence have also been documented. The aim of this study was to investigate the pathogenicity and potential mechanisms of virulence dependent on the tyrosine kinase BceF and phosphotyrosine phosphatase BceD of the cystic fibrosis opportunistic pathogen Burkholderia contaminans IST408. The insertion mutants bceD::Tp and bceF::Tp showed similar attenuation of adhesion and invasion of the cystic fibrosis lung epithelial cell line CFBE41o- compared to the parental strain B. contaminans IST408. In the absence of bceD or bceF genes, B. contaminans also showed a reduction in the ability to translocate across polarized epithelial cell monolayers, demonstrated by a higher transepithelial electrical resistance, reduced flux of fluorescein isothiocyanate-labeled bovine serum albumin, and higher levels of tight junction proteins ZO-1, occludin, and claudin-1 present in monolayers exposed to these bacterial mutants. Furthermore, bceD::Tp and bceF::Tp mutants induced lower levels of interleukin-6 (IL-6) and IL-8 release than the parental strain. In conclusion, although the mechanisms of pathogenicity dependent on BceD and BceF are not understood, these proteins contribute to the virulence of Burkholderia by enhancement of cell attachment and invasion, disruption of epithelial integrity, and modulation of the proinflammatory response.