title: The mgtC gene of Burkholderia cenocepacia is required for growth under magnesium limitation conditions and intracellular survival in macrophages.
abstract: Burkholderia cenocepacia, a bacterium commonly found in the environment, is an important opportunistic pathogen in patients with cystic fibrosis (CF). Very little is known about the mechanisms by which B. cenocepacia causes disease, but chronic infection of the airways in CF patients may be associated, at least in part, with the ability of this bacterium to survive within epithelial cells and macrophages. Survival in macrophages occurs in a membrane-bound compartment that is distinct from the lysosome, suggesting that B. cenocepacia prevents phagolysosomal fusion. In a previous study, we employed signature-tagged mutagenesis and an agar bead model of chronic pulmonary infection in rats to identify B. cenocepacia genes that are required for bacterial survival in vivo. One of the most significantly attenuated mutants had an insertion in the mgtC gene. Here, we show that mgtC is also needed for growth of B. cenocepacia in magnesium-depleted medium and for bacterial survival within murine macrophages. Using fluorescence microscopy, we demonstrated that B. cenocepacia mgtC mutants, unlike the parental isolate, colocalize with the fluorescent acidotropic probe LysoTracker Red. At 4 h postinfection, mgtC mutants expressing monomeric red fluorescent protein cannot retain this protein within the bacterial cytoplasm. Together, these results demonstrate that, unlike the parental strain, an mgtC mutant does not induce a delay in phagolysosomal fusion and the bacterium-containing vacuoles are rapidly targeted to the lysosome, where bacteria are destroyed.
authors.affiliation: Infectious Diseases Research Group, Siebens-Drake Research Institute, Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, N6A 5C1, Canada.
authors.forename: Kendra E
authors.initials: KE
authors.lastname: Maloney
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Valvano
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D042602:Burkholderia cepacia complex / Q000235:genetics* / Q000254:growth & development* / Q000648:ultrastructure; D027682:Cation Transport Proteins / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D004251:DNA Transposable Elements; D005798:Genes, Bacterial; D008264:Macrophages / Q000166:cytology / Q000382:microbiology*; D008274:Magnesium / Q000378:metabolism*; D051379:Mice; D016254:Mutagenesis, Insertional; D009154:Mutation; D026361:Reactive Nitrogen Species / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D027682:Cation Transport Proteins; D004251:DNA Transposable Elements; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; D008274:Magnesium