title: A Burkholderia cenocepacia gene encoding a non-functional tyrosine phosphatase is required for the delayed maturation of the bacteria-containing vacuoles in macrophages.
abstract: Burkholderia cenocepacia infects patients with cystic fibrosis. We have previously shown that B. cenocepacia can survive in macrophages within membrane vacuoles [B. cenocepacia-containing vacuoles (BcCVs)] that preclude fusion with the lysosome. The bacterial factors involved in B. cenocepacia intracellular survival are not fully elucidated. We report here that deletion of BCAM0628, encoding a predicted low molecular weight protein tyrosine phosphatase (LMW-PTP) that is restricted to B. cenocepacia strains of the transmissible ET-12 clone, accelerates the maturation of the BcCVs. Compared to the parental strain and deletion mutants in other LMW-PTPs that are widely conserved in Burkholderia species, a greater proportion of BcCVs containing the ΔBCAM0628 mutant were targeted to the lysosome. Accelerated BcCV maturation was not due to reduced intracellular viability since ΔBCAM0628 survived and replicated in macrophages similarly to the parental strain. Therefore, BCAM0628 was referred to as dpm (delayed phagosome maturation). We provide evidence that the Dpm protein is secreted during growth in vitro and upon macrophage infection. Dpm secretion requires an N-terminal signal peptide. Heterologous expression of Dpm in Burkholderia multivorans confers to this bacterium a similar phagosomal maturation delay to that found with B. cenocepacia. We demonstrate that Dpm is an inactive phosphatase, suggesting that its contribution to phagosomal maturation arrest must be unrelated to tyrosine phosphatase activity.
authors.affiliation: Centre for Human Immunology and Department of Microbiology and Immunology, University of Western Ontario, London, ON N6A 5C1, Canada.
authors.forename: Angel
authors.initials: A
authors.lastname: Andrade
authors.affiliation: Centre for Infection and Immunity, Queen's University Belfast, Belfast BT9 5AE, UK.|Centre for Human Immunology and Department of Microbiology and Immunology, University of Western Ontario, London, ON N6A 5C1, Canada.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Valvano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D057508:Burkholderia cenocepacia / Q000166:cytology / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D016680:Genome, Bacterial / Q000235:genetics*; D008264:Macrophages / Q000166:cytology / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010588:Phagosomes / Q000382:microbiology; D021382:Protein Sorting Signals; D017027:Protein Tyrosine Phosphatases / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014617:Vacuoles / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D017027:Protein Tyrosine Phosphatases