title: Mycobacterium tuberculosis phagosomes exhibit altered calmodulin-dependent signal transduction: contribution to inhibition of phagosome-lysosome fusion and intracellular survival in human macrophages.
abstract: Mycobacterium tuberculosis successfully parasitizes macrophages by disrupting the maturation of its phagosome, creating an intracellular compartment with endosomal rather than lysosomal characteristics. We have recently demonstrated that live M. tuberculosis infect human macrophages in the absence of an increase in cytosolic Ca(2+) ([Ca(2+)](c)), which correlates with inhibition of phagosome-lysosome fusion and intracellular viability. In contrast, killed M. tuberculosis induces an elevation in [Ca(2+)](c) that is coupled to phagosome-lysosome fusion. We tested the hypothesis that defective activation of the Ca(2+)-dependent effector proteins calmodulin (CaM) and CaM-dependent protein kinase II (CaMKII) contributes to the intracellular pathogenesis of tuberculosis. Phagosomes containing live M. tuberculosis exhibited decreased levels of CaM and the activated form of CaMKII compared with phagosomes encompassing killed tubercle bacilli. Furthermore, ionophore-induced elevations in [Ca(2+)](c) resulted in recruitment of CaM and activation of CaMKII on phagosomes containing live M. tuberculosis. Specific inhibitors of CaM or CaMKII blocked Ca(2+) ionophore-induced phagosomal maturation and enhanced the bacilli's intracellular viability. These results demonstrate a novel role for CaM and CaMKII in the regulation of phagosome-lysosome fusion and suggest that defective activation of these Ca(2+)-activated signaling components contributes to the successful parasitism of human macrophages by M. tuberculosis.
authors.affiliation: Inflammation Program, Graduate Program in Immunology, University of Iowa and Veterans Administration Medical Center, Iowa City, IA 52242, USA.
authors.forename: Z A
authors.initials: ZA
authors.lastname: Malik
authors.forename: S S
authors.initials: SS
authors.lastname: Iyer
authors.forename: D J
authors.initials: DJ
authors.lastname: Kusner
mesh_terms: D000328:Adult; D002118:Calcium / Q000037:antagonists & inhibitors / Q000502:physiology; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002147:Calmodulin / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D003600:Cytosol / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D004789:Enzyme Activation / Q000276:immunology; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D008247:Lysosomes / Q000276:immunology / Q000382:microbiology*; D008264:Macrophages / Q000201:enzymology / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D009169:Mycobacterium tuberculosis / Q000254:growth & development / Q000276:immunology* / Q000472:pathogenicity*; D010588:Phagosomes / Q000201:enzymology / Q000276:immunology / Q000382:microbiology*; D015398:Signal Transduction / Q000276:immunology*
chemical_list: D002147:Calmodulin; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002118:Calcium