Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text PDF (Full Text) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Fratti, R. A. Articles by Deretic, V. Search for Related Content PubMed PubMed Citation Articles by Fratti, R. A. Articles by Deretic, V. Related Collections Related Article Social Bookmarking                      What's this?

¹ Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109
² Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY 10461
³ Department of Biochemistry, University of Geneva, CH-1211 Geneva, Switzerland
⁴ Department of Molecular Medicine and Cell Biology, University of Massachusetts Medical School, Worcester, MA 01655
⁵ Program in Cellular and Molecular Biology, University of Michigan Medical School, Ann Arbor, MI 48109

Address correspondence to Vojo Deretic, University of Michigan Medical School, 5641 Medical Science Bldg. II, Ann Arbor, MI 48109-0620. Tel.: (734) 763-1580. Fax: (734) 647-6243. E-mail: deretic{at}umich.edu

Phagosomal biogenesis is a fundamental biological process of particular significance for the function of phagocytic and antigen-presenting cells. The precise mechanisms governing maturation of phagosomes into phagolysosomes are not completely understood. Here, we applied the property of pathogenic mycobacteria to cause phagosome maturation arrest in infected macrophages as a tool to dissect critical steps in phagosomal biogenesis. We report the requirement for 3-phosphoinositides and acquisition of Rab5 effector early endosome autoantigen (EEA1) as essential molecular events necessary for phagosomal maturation. Unlike the model phagosomes containing latex beads, which transiently recruited EEA1, mycobacterial phagosomes excluded this regulator of vesicular trafficking that controls membrane tethering and fusion processes within the endosomal pathway and is recruited to endosomal membranes via binding to phosphatidylinositol 3-phosphate (PtdIns[3]P). Inhibitors of phosphatidylinositol 3'(OH)-kinase (PI-3K) activity diminished EEA1 recruitment to newly formed latex bead phagosomes and blocked phagosomal acquisition of late endocytic properties, indicating that generation of PtdIns(3)P plays a role in phagosomal maturation. Microinjection into macrophages of antibodies against EEA1 and the PI-3K hVPS34 reduced acquisition of late endocytic markers by latex bead phagosomes, demonstrating an essential role of these Rab5 effectors in phagosomal biogenesis. The mechanism of EEA1 exclusion from mycobacterial phagosomes was investigated using mycobacterial products. Coating of latex beads with the major mycobacterial cell envelope glycosylated phosphatidylinositol lipoarabinomannan isolated from the virulent Mycobacterium tuberculosis H37Rv, inhibited recruitment of EEA1 to latex bead phagosomes, and diminished their maturation. These findings define the generation of phosphatidylinositol 3-phosphate and EEA1 recruitment as: (a) important regulatory events in phagosomal maturation and (b) critical molecular targets affected by M. tuberculosis. This study also identifies mycobacterial phosphoinositides as products with specialized toxic properties, interfering with discrete trafficking stages in phagosomal maturation.

Key Words: EEA1; endosome; hVPS34; LBPA; LAM

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

Related Article

How to survive a macrophage attack

Alan W. Dove
J. Cell Biol. 2001 154: 478. [Full Text] [PDF]

This article has been cited by other articles:

• Rampini, S. K., Selchow, P., Keller, C., Ehlers, S., Bottger, E. C., Sander, P. (2008). LspA inactivation in Mycobacterium tuberculosis results in attenuation without affecting phagosome maturation arrest. Microbiology 154: 2991-3001 [Abstract] [Full Text]  
• Philips, J. A., Porto, M. C., Wang, H., Rubin, E. J., Perrimon, N. (2008). ESCRT factors restrict mycobacterial growth. Proc. Natl. Acad. Sci. USA 105: 3070-3075 [Abstract] [Full Text]  
• Brumell, J. H., Scidmore, M. A. (2007). Manipulation of Rab GTPase Function by Intracellular Bacterial Pathogens. Microbiol. Mol. Biol. Rev. 71: 636-652 [Abstract] [Full Text]  
• Sun, J., Deghmane, A.-E., Soualhine, H., Hong, T., Bucci, C., Solodkin, A., Hmama, Z. (2007). Mycobacterium bovis BCG disrupts the interaction of Rab7 with RILP contributing to inhibition of phagosome maturation. J. Leukoc. Biol. 82: 1437-1445 [Abstract] [Full Text]  
• Huynh, K. K., Grinstein, S. (2007). Regulation of Vacuolar pH and Its Modulation by Some Microbial Species. Microbiol. Mol. Biol. Rev. 71: 452-462 [Abstract] [Full Text]  
• Herrada, A. A., Contreras, F. J., Tobar, J. A., Pacheco, R., Kalergis, A. M. (2007). Immune complex-induced enhancement of bacterial antigen presentation requires Fc{gamma} Receptor III expression on dendritic cells. Proc. Natl. Acad. Sci. USA 104: 13402-13407 [Abstract] [Full Text]  
• Fang, J., Brzostowski, J. A., Ou, S., Isik, N., Nair, V., Jin, T. (2007). A vesicle surface tyrosine kinase regulates phagosome maturation. J. Cell Biol. 178: 411-423 [Abstract] [Full Text]  
• MacGurn, J. A., Cox, J. S. (2007). A Genetic Screen for Mycobacterium tuberculosis Mutants Defective for Phagosome Maturation Arrest Identifies Components of the ESX-1 Secretion System. Infect. Immun. 75: 2668-2678 [Abstract] [Full Text]  
• Roberts, E. A., Chua, J., Kyei, G. B., Deretic, V. (2006). Higher order Rab programming in phagolysosome biogenesis. J. Cell Biol. 174: 923-929 [Abstract] [Full Text]  
• Fernandez-Moreira, E., Helbig, J. H., Swanson, M. S. (2006). Membrane Vesicles Shed by Legionella pneumophila Inhibit Fusion of Phagosomes with Lysosomes.. Infect. Immun. 74: 3285-3295 [Abstract] [Full Text]  
• Jarry, T. M., Cheung, A. L. (2006). Staphylococcus aureus Escapes More Efficiently from the Phagosome of a Cystic Fibrosis Bronchial Epithelial Cell Line than from Its Normal Counterpart.. Infect. Immun. 74: 2568-2577 [Abstract] [Full Text]  
• D'Avila, H., Melo, R. C. N., Parreira, G. G., Werneck-Barroso, E., Castro-Faria-Neto, H. C., Bozza, P. T. (2006). Mycobacterium bovis Bacillus Calmette-Guerin Induces TLR2-Mediated Formation of Lipid Bodies: Intracellular Domains for Eicosanoid Synthesis In Vivo.. J. Immunol. 176: 3087-3097 [Abstract] [Full Text]  
• Lindmo, K., Stenmark, H. (2006). Regulation of membrane traffic by phosphoinositide 3-kinases. J. Cell Sci. 119: 605-614 [Abstract] [Full Text]  
• Lang, M. L., Glatman-Freedman, A. (2006). Do CD1-Restricted T Cells Contribute to Antibody-Mediated Immunity against Mycobacterium tuberculosis?. Infect. Immun. 74: 803-809 [Full Text]  
• Toyooka, K., Takai, S., Kirikae, T. (2005). Rhodococcus equi can survive a phagolysosomal environment in macrophages by suppressing acidification of the phagolysosome. J Med Microbiol 54: 1007-1015 [Abstract] [Full Text]  
• Byfield, M. P., Murray, J. T., Backer, J. M. (2005). hVps34 Is a Nutrient-regulated Lipid Kinase Required for Activation of p70 S6 Kinase. J. Biol. Chem. 280: 33076-33082 [Abstract] [Full Text]  
• Sendide, K., Reiner, N. E., Lee, J. S. I., Bourgoin, S., Talal, A., Hmama, Z. (2005). Cross-Talk between CD14 and Complement Receptor 3 Promotes Phagocytosis of Mycobacteria: Regulation by Phosphatidylinositol 3-Kinase and Cytohesin-1. J. Immunol. 174: 4210-4219 [Abstract] [Full Text]  
• Walch, M., Eppler, E., Dumrese, C., Barman, H., Groscurth, P., Ziegler, U. (2005). Uptake of Granulysin via Lipid Rafts Leads to Lysis of Intracellular Listeria innocua. J. Immunol. 174: 4220-4227 [Abstract] [Full Text]  
• Lee, W. L., Kim, M.-K., Schreiber, A. D., Grinstein, S. (2005). Role of Ubiquitin and Proteasomes in Phagosome Maturation. Mol. Biol. Cell 16: 2077-2090 [Abstract] [Full Text]  
• Shohdy, N., Efe, J. A., Emr, S. D., Shuman, H. A. (2005). Pathogen effector protein screening in yeast identifies Legionella factors that interfere with membrane trafficking. Proc. Natl. Acad. Sci. USA 102: 4866-4871 [Abstract] [Full Text]  
• Thompson, C. R., Iyer, S. S., Melrose, N., VanOosten, R., Johnson, K., Pitson, S. M., Obeid, L. M., Kusner, D. J. (2005). Sphingosine Kinase 1 (SK1) Is Recruited to Nascent Phagosomes in Human Macrophages: Inhibition of SK1 Translocation by Mycobacterium tuberculosis. J. Immunol. 174: 3551-3561 [Abstract] [Full Text]  
• Vergne, I., Chua, J., Lee, H.-H., Lucas, M., Belisle, J., Deretic, V. (2005). Mechanism of phagolysosome biogenesis block by viable Mycobacterium tuberculosis. Proc. Natl. Acad. Sci. USA 102: 4033-4038 [Abstract] [Full Text]  
• Ramachandra, L., Smialek, J. L., Shank, S. S., Convery, M., Boom, W. H., Harding, C. V. (2005). Phagosomal Processing of Mycobacterium tuberculosis Antigen 85B Is Modulated Independently of Mycobacterial Viability and Phagosome Maturation. Infect. Immun. 73: 1097-1105 [Abstract] [Full Text]  
• Fratti, R. A., Jun, Y., Merz, A. J., Margolis, N., Wickner, W. (2004). Interdependent assembly of specific regulatory lipids and membrane fusion proteins into the vertex ring domain of docked vacuoles. J. Cell Biol. 167: 1087-1098 [Abstract] [Full Text]  
• Koo, H. C., Park, Y. H., Hamilton, M. J., Barrington, G. M., Davies, C. J., Kim, J. B., Dahl, J. L., Waters, W. R., Davis, W. C. (2004). Analysis of the Immune Response to Mycobacterium avium subsp. paratuberculosis in Experimentally Infected Calves. Infect. Immun. 72: 6870-6883 [Abstract] [Full Text]  
• Swanson, J. A., Hoppe, A. D. (2004). The coordination of signaling during Fc receptor-mediated phagocytosis. J. Leukoc. Biol. 76: 1093-1103 [Abstract] [Full Text]  
• Saito-Nakano, Y., Yasuda, T., Nakada-Tsukui, K., Leippe, M., Nozaki, T. (2004). Rab5-associated Vacuoles Play a Unique Role in Phagocytosis of the Enteric Protozoan Parasite Entamoeba histolytica. J. Biol. Chem. 279: 49497-49507 [Abstract] [Full Text]  
• Pethe, K., Swenson, D. L., Alonso, S., Anderson, J., Wang, C., Russell, D. G. (2004). Isolation of Mycobacterium tuberculosis mutants defective in the arrest of phagosome maturation. Proc. Natl. Acad. Sci. USA 101: 13642-13647 [Abstract] [Full Text]  
• Firoved, A. M., Ornatowski, W., Deretic, V. (2004). Microarray Analysis Reveals Induction of Lipoprotein Genes in Mucoid Pseudomonas aeruginosa: Implications for Inflammation in Cystic Fibrosis. Infect. Immun. 72: 5012-5018 [Abstract] [Full Text]  
• Chua, J., Deretic, V. (2004). Mycobacterium tuberculosis Reprograms Waves of Phosphatidylinositol 3-Phosphate on Phagosomal Organelles. J. Biol. Chem. 279: 36982-36992 [Abstract] [Full Text]  
• Drecktrah, D., Knodler, L. A., Steele-Mortimer, O. (2004). Modulation and Utilization of Host Cell Phosphoinositides by Salmonella spp.. Infect. Immun. 72: 4331-4335 [Full Text]  
• Walburger, A., Koul, A., Ferrari, G., Nguyen, L., Prescianotto-Baschong, C., Huygen, K., Klebl, B., Thompson, C., Bacher, G., Pieters, J. (2004). Protein Kinase G from Pathogenic Mycobacteria Promotes Survival Within Macrophages. Science 304: 1800-1804 [Abstract] [Full Text]  
• Vieira, O. V., Harrison, R. E., Scott, C. C., Stenmark, H., Alexander, D., Liu, J., Gruenberg, J., Schreiber, A. D., Grinstein, S. (2004). Acquisition of Hrs, an Essential Component of Phagosomal Maturation, Is Impaired by Mycobacteria. Mol. Cell. Biol. 24: 4593-4604 [Abstract] [Full Text]  
• Miller, B. H., Fratti, R. A., Poschet, J. F., Timmins, G. S., Master, S. S., Burgos, M., Marletta, M. A., Deretic, V. (2004). Mycobacteria Inhibit Nitric Oxide Synthase Recruitment to Phagosomes during Macrophage Infection. Infect. Immun. 72: 2872-2878 [Abstract] [Full Text]  
• Alexander, D. C., Jones, J. R. W., Tan, T., Chen, J. M., Liu, J. (2004). PimF, a Mannosyltransferase of Mycobacteria, Is Involved in the Biosynthesis of Phosphatidylinositol Mannosides and Lipoarabinomannan. J. Biol. Chem. 279: 18824-18833 [Abstract] [Full Text]  
• Hmama, Z., Sendide, K., Talal, A., Garcia, R., Dobos, K., Reiner, N. E. (2004). Quantitative analysis of phagolysosome fusion in intact cells: inhibition by mycobacterial lipoarabinomannan and rescue by an 1{alpha},25-dihydroxyvitamin D3-phosphoinositide 3-kinase pathway. J. Cell Sci. 117: 2131-2140 [Abstract] [Full Text]  
• Lee, J. S., Hmama, Z., Mui, A., Reiner, N. E. (2004). Stable Gene Silencing in Human Monocytic Cell Lines Using Lentiviral-delivered Small Interference RNA: SILENCING OF THE p110{alpha} ISOFORM OF PHOSPHOINOSITIDE 3-KINASE REVEALS DIFFERENTIAL REGULATION OF ADHERENCE INDUCED BY 1{alpha},25-DIHYDROXYCHOLECALCIFEROL AND BACTERIAL LIPOPOLYSACCHARIDE. J. Biol. Chem. 279: 9379-9388 [Abstract] [Full Text]  
• Vergne, I., Fratti, R. A., Hill, P. J., Chua, J., Belisle, J., Deretic, V. (2004). Mycobacterium tuberculosis Phagosome Maturation Arrest: Mycobacterial Phosphatidylinositol Analog Phosphatidylinositol Mannoside Stimulates Early Endosomal Fusion. Mol. Biol. Cell 15: 751-760 [Abstract] [Full Text]  
• Fratti, R. A., Chua, J., Deretic, V. (2003). Induction of p38 Mitogen-activated Protein Kinase Reduces Early Endosome Autoantigen 1 (EEA1) Recruitment to Phagosomal Membranes. J. Biol. Chem. 278: 46961-46967 [Abstract] [Full Text]  
• Harrison, R. E., Bucci, C., Vieira, O. V., Schroer, T. A., Grinstein, S. (2003). Phagosomes Fuse with Late Endosomes and/or Lysosomes by Extension of Membrane Protrusions along Microtubules: Role of Rab7 and RILP. Mol. Cell. Biol. 23: 6494-6506 [Abstract] [Full Text]  
• Woolsey, A. M., Sunwoo, L., Petersen, C. A., Brachmann, S. M., Cantley, L. C., Burleigh, B. A. (2003). Novel PI 3-kinase-dependent mechanisms of trypanosome invasion and vacuole maturation. J. Cell Sci. 116: 3611-3622 [Abstract] [Full Text]  
• Vergne, I., Chua, J., Deretic, V. (2003). Tuberculosis Toxin Blocking Phagosome Maturation Inhibits a Novel Ca2+/Calmodulin-PI3K hVPS34 Cascade. J. Exp. Med. 198: 653-659 [Abstract] [Full Text]  
• Indrigo, J., Hunter, R. L. Jr, Actor, J. K. (2003). Cord factor trehalose 6,6'-dimycolate (TDM) mediates trafficking events during mycobacterial infection of murine macrophages. Microbiology 149: 2049-2059 [Abstract] [Full Text]  
• Kelley, V. A., Schorey, J. S. (2003). Mycobacterium's Arrest of Phagosome Maturation in Macrophages Requires Rab5 Activity and Accessibility to Iron. Mol. Biol. Cell 14: 3366-3377 [Abstract] [Full Text]  
• Fratti, R. A., Chua, J., Vergne, I., Deretic, V. (2003). Mycobacterium tuberculosis glycosylated phosphatidylinositol causes phagosome maturation arrest. Proc. Natl. Acad. Sci. USA 100: 5437-5442 [Abstract] [Full Text]  
• Booth, J. W., Telio, D., Liao, E. H., McCaw, S. E., Matsuo, T., Grinstein, S., Gray-Owen, S. D. (2003). Phosphatidylinositol 3-Kinases in Carcinoembryonic Antigen-related Cellular Adhesion Molecule-mediated Internalization of Neisseria gonorrhoeae. J. Biol. Chem. 278: 14037-14045 [Abstract] [Full Text]  
• Vieira, O. V., Bucci, C., Harrison, R. E., Trimble, W. S., Lanzetti, L., Gruenberg, J., Schreiber, A. D., Stahl, P. D., Grinstein, S. (2003). Modulation of Rab5 and Rab7 Recruitment to Phagosomes by Phosphatidylinositol 3-Kinase. Mol. Cell. Biol. 23: 2501-2514 [Abstract] [Full Text]  
• Windmiller, D. A., Backer, J. M. (2003). Distinct Phosphoinositide 3-Kinases Mediate Mast Cell Degranulation in Response to G-protein-coupled Versus Fcepsilon RI Receptors. J. Biol. Chem. 278: 11874-11878 [Abstract] [Full Text]  
• Malik, Z. A., Thompson, C. R., Hashimi, S., Porter, B., Iyer, S. S., Kusner, D. J. (2003). Cutting Edge: Mycobacterium tuberculosis Blocks Ca2+ Signaling and Phagosome Maturation in Human Macrophages Via Specific Inhibition of Sphingosine Kinase. J. Immunol. 170: 2811-2815 [Abstract] [Full Text]  
• Ibrahim-Granet, O., Philippe, B., Boleti, H., Boisvieux-Ulrich, E., Grenet, D., Stern, M., Latge, J. P. (2003). Phagocytosis and Intracellular Fate of Aspergillus fumigatus Conidia in Alveolar Macrophages. Infect. Immun. 71: 891-903 [Abstract] [Full Text]  
• Garcia-Garcia, E., Rosales, C. (2002). Signal transduction during Fc receptor-mediated phagocytosis. J. Leukoc. Biol. 72: 1092-1108 [Abstract] [Full Text]  
• Mukherjee, K., Parashuraman, S., Krishnamurthy, G., Majumdar, J., Yadav, A., Kumar, R., Basu, S. K., Mukhopadhyay, A. (2002). Diverting intracellular trafficking of Salmonella to the lysosome through activation of the late endocytic Rab7 by intracellular delivery of muramyl dipeptide. J. Cell Sci. 115: 3693-3701 [Abstract] [Full Text]  
• Russell, D. G., Mwandumba, H. C., Rhoades, E. E. (2002). Mycobacterium and the coat of many lipids. J. Cell Biol. 158: 421-426 [Abstract] [Full Text]  
• Ishii, K. J., Takeshita, F., Gursel, I., Gursel, M., Conover, J., Nussenzweig, A., Klinman, D. M. (2002). Potential Role of Phosphatidylinositol 3 Kinase, rather than DNA-dependent Protein Kinase, in CpG DNA-induced Immune Activation. J. Exp. Med. 196: 269-274 [Abstract] [Full Text]  
• Fratti, R. A., Chua, J., Deretic, V. (2002). Cellubrevin Alterations and Mycobacterium tuberculosis Phagosome Maturation Arrest. J. Biol. Chem. 277: 17320-17326 [Abstract] [Full Text]  
• Perskvist, N., Roberg, K., Kulyte, A., Stendahl, O. (2002). Rab5a GTPase regulates fusion between pathogen-containing phagosomes and cytoplasmic organelles in human neutrophils. J. Cell Sci. 115: 1321-1330 [Abstract] [Full Text]  
• Gillooly, D. J., Simonsen, A., Stenmark, H. (2001). Phosphoinositides and phagocytosis. J. Cell Biol. 155: 15-18 [Abstract] [Full Text]  
• Vieira, O. V., Botelho, R. J., Rameh, L., Brachmann, S. M., Matsuo, T., Davidson, H. W., Schreiber, A., Backer, J. M., Cantley, L. C., Grinstein, S. (2001). Distinct roles of class I and class III phosphatidylinositol 3-kinases in phagosome formation and maturation. J. Cell Biol. 155: 19-26 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents