title: Tyrosine Phosphorylation and Dephosphorylation in Burkholderia cenocepacia Affect Biofilm Formation, Growth under Nutritional Deprivation, and Pathogenicity.
abstract: Burkholderia cenocepacia, a member of the B. cepacia complex (Bcc), is an opportunistic pathogen causing serious chronic infections in patients with cystic fibrosis. Tyrosine phosphorylation has emerged as an important posttranslational modification modulating the physiology and pathogenicity of Bcc bacteria. Here, we investigated the predicted bacterial tyrosine kinases BCAM1331 and BceF and the low-molecular-weight protein tyrosine phosphatases BCAM0208, BceD, and BCAL2200 of B. cenocepacia K56-2. We show that BCAM1331, BceF, BCAM0208, and BceD contribute to biofilm formation, while BCAL2200 is required for growth under nutrient-limited conditions. Multiple deletions of either tyrosine kinase or low-molecular-weight protein tyrosine phosphatase genes resulted in the attenuation of B. cenocepacia intramacrophage survival and reduced pathogenicity in the Galleria mellonella larval infection model. Experimental evidence indicates that BCAM1331 displays reduced tyrosine autophosphorylation activity compared to that of BceF. With the artificial substrate p-nitrophenyl phosphate, the phosphatase activities of the three low-molecular-weight protein tyrosine phosphatases demonstrated similar kinetic parameters. However, only BCAM0208 and BceD could dephosphorylate BceF. Further, BCAL2200 became tyrosine phosphorylated in vivo and catalyzed its autodephosphorylation. Together, our data suggest that despite having similar biochemical activities, low-molecular-weight protein tyrosine phosphatases and tyrosine kinases have both overlapping and specific roles in the physiology of B. cenocepacia.
authors.affiliation: Centre for Human Immunology and Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada Departamento de Microbiología, Facultad de Medicina, Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, México.
authors.forename: Angel
authors.initials: A
authors.lastname: Andrade
authors.affiliation: Centre for Human Immunology and Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada Instituto de Biotecnología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Nuevo León, México.
authors.forename: Faviola
authors.initials: F
authors.lastname: Tavares-Carreón
authors.affiliation: Centre for Human Immunology and Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada.
authors.forename: Maryam
authors.initials: M
authors.lastname: Khodai-Kalaki
authors.affiliation: Centre for Human Immunology and Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada Centre for Infection and Immunity, Queen's University Belfast, Belfast, United Kingdom m.valvano@qub.ac.uk.
authors.forename: Miguel A
authors.identifier: http://orcid.org/0000-0001-8229-3641
authors.initials: MA
authors.lastname: Valvano
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D018441:Biofilms / Q000254:growth & development*; D057508:Burkholderia cenocepacia / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D007814:Larva / Q000382:microbiology; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D009036:Moths / Q000382:microbiology; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D000067996:RAW 264.7 Cells; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases; D017027:Protein Tyrosine Phosphatases