title: 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.
abstract: 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.
authors.affiliation: Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
authors.forename: Ana S
authors.initials: AS
authors.lastname: Ferreira
authors.affiliation: Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
authors.forename: Inês N
authors.initials: IN
authors.lastname: Silva
authors.affiliation: Centro de Química Física Molecular and Institute of Nanoscience and Nanotechnology, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
authors.forename: Fábio
authors.initials: F
authors.lastname: Fernandes
authors.affiliation: Centre of Microbial Host Interactions, Institute of Technology Tallaght, Dublin, Ireland.
authors.forename: Ruth
authors.initials: R
authors.lastname: Pilkington
authors.affiliation: Centre of Microbial Host Interactions, Institute of Technology Tallaght, Dublin, Ireland.
authors.forename: Máire
authors.initials: M
authors.lastname: Callaghan
authors.affiliation: Centre of Microbial Host Interactions, Institute of Technology Tallaght, Dublin, Ireland.
authors.forename: Siobhán
authors.initials: S
authors.lastname: McClean
authors.affiliation: Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal lmoreira@tecnico.ulisboa.pt.
authors.forename: Leonilde M
authors.identifier: http://orcid.org/0000-0002-6838-4245
authors.initials: LM
authors.lastname: Moreira
mesh_terms: D000418:Albumins / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001422:Bacterial Adhesion; D019121:Burkholderia Infections / Q000382:microbiology / Q000473:pathology; D042602:Burkholderia cepacia complex / Q000201:enzymology / Q000235:genetics / Q000472:pathogenicity*; D002460:Cell Line; D002939:Ciprofloxacin / Q000494:pharmacology; D062445:Claudin-1 / Q000096:biosynthesis; D003550:Cystic Fibrosis / Q000382:microbiology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D017097:Electric Impedance; D006801:Humans; D007249:Inflammation / Q000276:immunology; D015850:Interleukin-6 / Q000096:biosynthesis / Q000378:metabolism; D016209:Interleukin-8 / Q000096:biosynthesis / Q000378:metabolism; D008168:Lung / Q000382:microbiology*; D008564:Membrane Potentials; D009154:Mutation; D062793:Occludin / Q000096:biosynthesis; D021381:Protein Transport; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000502:physiology*; D020545:Respiratory Mucosa / Q000382:microbiology*; D019108:Tight Junctions / Q000473:pathology; D037521:Virulence Factors / Q000235:genetics*; D062826:Zonula Occludens-1 Protein / Q000096:biosynthesis
chemical_list: D000418:Albumins; D000900:Anti-Bacterial Agents; C505032:CFTR protein, human; C508606:CXCL8 protein, human; D062445:Claudin-1; C508600:IL6 protein, human; D015850:Interleukin-6; D016209:Interleukin-8; D062793:Occludin; C569042:TJP1 protein, human; D037521:Virulence Factors; D062826:Zonula Occludens-1 Protein; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D002939:Ciprofloxacin; D011505:Protein-Tyrosine Kinases; D017027:Protein Tyrosine Phosphatases