Mycobacterium tuberculosis protein tyrosine phosphatase (PtpA) excludes host vacuolar-H+–ATPase to inhibit phagosome acidification

Mycobacterium tuberculosis (Mtb) pathogenicity depends on its ability to inhibit phagosome acidification and maturation processes after engulfment by macrophages. Here, we show that the secreted Mtb protein tyrosine phosphatase (PtpA) binds to subunit H of the macrophage vacuolar-H+-ATPase (V-ATPase) machinery, a multisubunit protein complex in the phagosome membrane that drives luminal acidification. Furthermore, we show that the macrophage class C vacuolar protein sorting complex, a key regulator of endosomal membrane fusion, associates with V-ATPase in phagosome maturation, suggesting a unique role for V-ATPase in coordinating phagosome–lysosome fusion. PtpA interaction with host V-ATPase is required for the previously reported dephosphorylation of VPS33B and subsequent exclusion of V-ATPase from the phagosome during Mtb infection. These findings show that inhibition of phagosome acidification in the mycobacterial phagosome is directly attributed to PtpA, a key protein needed for Mtb survival and pathogenicity within host macrophages.

[1]  C. Brett,et al.  Subunit organization and Rab interactions of Vps-C protein complexes that control endolysosomal membrane traffic , 2011, Molecular biology of the cell.

[2]  N. Hacohen,et al.  Mycobacterium tuberculosis protein ESAT‐6 is a potent activator of the NLRP3/ASC inflammasome , 2010, Cellular microbiology.

[3]  K. Papavinasasundaram,et al.  Convergence of Ser/Thr and Two-component Signaling to Coordinate Expression of the Dormancy Regulon in Mycobacterium tuberculosis* , 2010, The Journal of Biological Chemistry.

[4]  C. Dye,et al.  The Population Dynamics and Control of Tuberculosis , 2010, Science.

[5]  R. Carano,et al.  Host-Detrimental Role of Esx-1-Mediated Inflammasome Activation in Mycobacterial Infection , 2010, PLoS pathogens.

[6]  H. Bach,et al.  Protein kinase and phosphatase signaling in Mycobacterium tuberculosis physiology and pathogenesis. , 2010, Biochimica et biophysica acta.

[7]  M. Swanson,et al.  Inhibition of Host Vacuolar H+-ATPase Activity by a Legionella pneumophila Effector , 2010, PLoS pathogens.

[8]  J. Bernardo,et al.  Initial cytoplasmic and phagosomal consequences of human neutrophil exposure to Staphylococcus epidermidis , 2009, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[9]  C. Brett,et al.  Vps-c Complexes: Gatekeepers of Endolysosomal Traffic This Review Comes from a Themed Issue on Membranes and Organelles Edited Vps-c Complexes: Molecular Organization Cellular Functions and Regulation , 2022 .

[10]  Nobuyuki Kawamura,et al.  Direct recruitment of H+-ATPase from lysosomes for phagosomal acidification , 2009, Journal of Cell Science.

[11]  H. Bach,et al.  Mycobacterium tuberculosis PtkA is a novel protein tyrosine kinase whose substrate is PtpA. , 2009, The Biochemical journal.

[12]  H. Bach,et al.  Mycobacterium tuberculosis virulence is mediated by PtpA dephosphorylation of human vacuolar protein sorting 33B. , 2008, Cell host & microbe.

[13]  A. Juillerat,et al.  A split-protein sensor for studying protein-protein interaction in mycobacteria. , 2008, Journal of microbiological methods.

[14]  Michael J. E. Sternberg,et al.  3D-Garden: a system for modelling protein-protein complexes based on conformational refinement of ensembles generated with the marching cubes algorithm , 2008, Bioinform..

[15]  O. Jensen,et al.  The vacuolar V1/V0‐ATPase is involved in the release of the HOPS subunit Vps41 from vacuoles, vacuole fragmentation and fusion , 2008, FEBS letters.

[16]  C. Bucci,et al.  Mycobacterium bovis BCG disrupts the interaction of Rab7 with RILP contributing to inhibition of phagosome maturation , 2007, Journal of leukocyte biology.

[17]  Michael Forgac,et al.  Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology , 2007, Nature Reviews Molecular Cell Biology.

[18]  C. Ostrowicz,et al.  The CORVET tethering complex interacts with the yeast Rab5 homolog Vps21 and is involved in endo-lysosomal biogenesis. , 2007, Developmental cell.

[19]  Samuel I. Miller,et al.  Activation of the bacterial sensor kinase PhoQ by acidic pH. , 2007, Molecular cell.

[20]  H. Bach,et al.  Mycobacterium avium subsp. paratuberculosis PtpA Is an Endogenous Tyrosine Phosphatase Secreted during Infection , 2006, Infection and Immunity.

[21]  Y. Av‐Gay,et al.  Mycobacterial manipulation of the host cell. , 2005, FEMS microbiology reviews.

[22]  I. Jutras,et al.  Phagocytosis: at the crossroads of innate and adaptive immunity. , 2005, Annual review of cell and developmental biology.

[23]  Sunil Q. Mehta,et al.  The v-ATPase V0 Subunit a1 Is Required for a Late Step in Synaptic Vesicle Exocytosis in Drosophila , 2005, Cell.

[24]  J. Luzio,et al.  Endocytic Delivery to Lysosomes Mediated by Concurrent Fusion and Kissing Events in Living Cells , 2005, Current Biology.

[25]  Y. Av‐Gay,et al.  Kinome Analysis of Host Response to Mycobacterial Infection: a Novel Technique in Proteomics , 2003, Infection and Immunity.

[26]  E. Alexander,et al.  Syntaxin Isoform Specificity in the Regulation of Renal H+-ATPase Exocytosis* , 2003, Journal of Biological Chemistry.

[27]  B. Peterlin,et al.  Subunit H of the V-ATPase Binds to the Medium Chain of Adaptor Protein Complex 2 and Connects Nef to the Endocytic Machinery* , 2002, The Journal of Biological Chemistry.

[28]  Y. Av‐Gay,et al.  Expression and localization of the Mycobacterium tuberculosis protein tyrosine phosphatase PtpA. , 2002, Research in microbiology.

[29]  Brian W. Matthews,et al.  Crystal structure of the regulatory subunit H of the V-type ATPase of Saccharomyces cerevisiae , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Scott D. Emr,et al.  New Component of the Vacuolar Class C-Vps Complex Couples Nucleotide Exchange on the Ypt7 Gtpase to Snare-Dependent Docking and Fusion , 2000, The Journal of cell biology.

[31]  Y Av-Gay,et al.  The eukaryotic-like Ser/Thr protein kinases of Mycobacterium tuberculosis. , 2000, Trends in microbiology.

[32]  T. Tjelle,et al.  Phagosome dynamics and function , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[33]  J. Condeelis,et al.  Mycobacterial infection of macrophages results in membrane-permeable phagosomes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[34]  H. Izumiya,et al.  Yersinia pseudotuberculosis blocks the phagosomal acidification of B10.A mouse macrophages through the inhibition of vacuolar H(+)-ATPase activity. , 1999, Microbial pathogenesis.

[35]  P. Constant,et al.  Phagosomal pH determination by dual fluorescence flow cytometry. , 1998, Analytical biochemistry.

[36]  S. Grinstein,et al.  Regulation of Phagosomal Acidification , 1997, The Journal of Biological Chemistry.

[37]  S. Emr,et al.  A novel RING finger protein complex essential for a late step in protein transport to the yeast vacuole. , 1997, Molecular biology of the cell.

[38]  M. Horwitz,et al.  The Mycobacterium tuberculosis phagosome interacts with early endosomes and is accessible to exogenously administered transferrin , 1996, The Journal of experimental medicine.

[39]  K. Sandvig,et al.  Inhibition of the vacuolar H(+)-ATPase with bafilomycin reduces delivery of internalized molecules from mature multivesicular endosomes to lysosomes in HEp-2 cells. , 1996, European journal of cell biology.

[40]  P. Schlesinger,et al.  Lack of acidification in Mycobacterium phagosomes produced by exclusion of the vesicular proton-ATPase. , 1994, Science.