Legionella Subvert the Functions of Rab1 and Sec22b to Create a Replicative Organelle

Legionella pneumophila is a bacterial pathogen that infects eukaryotic host cells and replicates inside a specialized organelle that is morphologically similar to the endoplasmic reticulum (ER). To better understand the molecular mechanisms governing transport of the Legionella-containing vacuole (LCV), we have identified host proteins that participate in the conversion of the LCV into a replicative organelle. Our data show that Rab1 is recruited to the LCV within minutes of uptake. Rab1 recruitment to the LCV precedes remodeling of this compartment by ER-derived vesicles. Genetic inhibition studies demonstrate that Rab1 is important for the recruitment of ER-derived vesicles to the LCV and that inhibiting Rab1 function abrogates intracellular growth of Legionella. Morphological studies indicate that the Sec22b protein is located on ER-derived vesicles recruited to the LCV and that Sec22b is delivered to the LCV membrane. Sec22b function was found to be important for biogenesis of the specialized organelle that supports Legionella replication. These studies demonstrate that Legionella has the ability to subvert Rab1 and Sec22b function to facilitate the transport and fusion of ER-derived vesicles with the LCV, resulting in the formation of a specialized organelle that can support bacterial replication.

[1]  Zhao-Qing Luo,et al.  Multiple substrates of the Legionella pneumophila Dot/Icm system identified by interbacterial protein transfer. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Randy Schekman,et al.  Multiple Cargo Binding Sites on the COPII Subunit Sec24p Ensure Capture of Diverse Membrane Proteins into Transport Vesicles , 2003, Cell.

[3]  E. Mossessova,et al.  SNARE Selectivity of the COPII Coat , 2003, Cell.

[4]  Charles Barlowe,et al.  Signals for COPII-dependent export from the ER: what's the ticket out? , 2003, Trends in cell biology.

[5]  G. Warren,et al.  Golgin‐84 is a rab1 Binding Partner Involved in Golgi Structure , 2003, Traffic.

[6]  V. Haucke.  Vesicle budding: a coat for the COPs. , 2003, Trends in cell biology.

[7]  C. Roy,et al.  Legionella phagosomes intercept vesicular traffic from endoplasmic reticulum exit sites , 2002, Nature Cell Biology.

[8]  R. Schekman,et al.  Cargo selection into COPII vesicles is driven by the Sec24p subunit , 2002, The EMBO journal.

[9]  C. Barlowe.  Three-dimensional structure of a COPII prebudding complex. , 2002, Developmental cell.

[10]  R. A. Corpina,et al.  Structure of the Sec23/24–Sar1 pre-budding complex of the COPII vesicle coat , 2002, Nature.

[11]  Etienne Gagnon,et al.  Endoplasmic Reticulum-Mediated Phagocytosis Is a Mechanism of Entry into Macrophages , 2002, Cell.

[12]  Bor Luen Tang,et al.  Early/recycling endosomes-to-TGN transport involves two SNARE complexes and a Rab6 isoform , 2002, The Journal of cell biology.

[13]  R. Kahn,et al.  A Bacterial Guanine Nucleotide Exchange Factor Activates ARF on Legionella Phagosomes , 2002, Science.

[14]  C. G. Robinson,et al.  How the parasitic bacterium Legionella pneumophila modifies its phagosome and transforms it into rough ER: implications for conversion of plasma membrane to the ER membrane. , 2001, Journal of cell science.

[15]  J. Lippincott-Schwartz,et al.  Maintenance of Golgi structure and function depends on the integrity of ER export , 2001, The Journal of cell biology.

[16]  W. Balch,et al.  Rab1 Interaction with a GM130 Effector Complex Regulates COPII Vesicle cis‐Golgi Tethering , 2001, Traffic.

[17]  J. Hay,et al.  Subunit Structure of a Mammalian ER/Golgi SNARE Complex* , 2000, The Journal of Biological Chemistry.

[18]  H. Nagai,et al.  Identification of Icm protein complexes that play distinct roles in the biogenesis of an organelle permissive for Legionella pneumophila intracellular growth , 2000, Molecular microbiology.

[19]  J. Rothman,et al.  Compartmental specificity of cellular membrane fusion encoded in SNARE proteins , 2000, Nature.

[20]  W. Balch,et al.  Rab1 recruitment of p115 into a cis-SNARE complex: programming budding COPII vesicles for fusion. , 2000, Science.

[21]  M. Horwitz,et al.  Deviant Expression of Rab5 on Phagosomes Containing the Intracellular Pathogens Mycobacterium tuberculosis andLegionella pneumophila Is Associated with Altered Phagosomal Fate , 2000, Infection and Immunity.

[22]  Ludger Johannes,et al.  Rab6 Coordinates a Novel Golgi to ER Retrograde Transport Pathway in Live Cells , 1999, The Journal of cell biology.

[23]  R. Pepperkok,et al.  Evidence for a COP-I-independent transport route from the Golgi complex to the endoplasmic reticulum , 1999, Nature Cell Biology.

[24]  C. Roy,et al.  Pore‐forming activity is not sufficient for Legionella pneumophila phagosome trafficking and intracellular growth , 1999, Molecular microbiology.

[25]  E. Tisdale.  A Rab2 mutant with impaired GTPase activity stimulates vesicle formation from pre-Golgi intermediates. , 1999, Molecular biology of the cell.

[26]  S. Wong,et al.  Morphological and functional association of Sec22b/ERS-24 with the pre-Golgi intermediate compartment. , 1999, Molecular biology of the cell.

[27]  F. Maxfield,et al.  Early Events in Phagosome Establishment Are Required for Intracellular Survival of Legionella pneumophila , 1998, Infection and Immunity.

[28]  J. Lippincott-Schwartz,et al.  Building a secretory apparatus: role of ARF1/COPI in Golgi biogenesis and maintenance , 1998, Histochemistry and Cell Biology.

[29]  R. Isberg,et al.  Legionella pneumophila DotA protein is required for early phagosome trafficking decisions that occur within minutes of bacterial uptake , 1998, Molecular microbiology.

[30]  H. Shuman,et al.  Host cell killing and bacterial conjugation require overlapping sets of genes within a 22-kb region of the Legionella pneumophila genome. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[31]  R. Isberg,et al.  Conjugative transfer by the virulence system of Legionella pneumophila. , 1998, Science.

[32]  Rainer Pepperkok,et al.  Visualization of ER-to-Golgi Transport in Living Cells Reveals a Sequential Mode of Action for COPII and COPI , 1997, Cell.

[33]  R. Scheller,et al.  Protein Interactions Regulating Vesicle Transport between the Endoplasmic Reticulum and Golgi Apparatus in Mammalian Cells , 1997, Cell.

[34]  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.

[35]  R. Naviaux,et al.  The pCL vector system: rapid production of helper-free, high-titer, recombinant retroviruses , 1996, Journal of virology.

[36]  W. Balch,et al.  Mammalian Sly1 Regulates Syntaxin 5 Function in Endoplasmic Reticulum to Golgi Transport* , 1996, The Journal of Biological Chemistry.

[37]  W. Balch,et al.  Sequential coupling between COPII and COPI vesicle coats in endoplasmic reticulum to Golgi transport , 1995, The Journal of cell biology.

[38]  M. Swanson,et al.  Association of Legionella pneumophila with the macrophage endoplasmic reticulum , 1995, Infection and immunity.

[39]  C. Barlowe,et al.  COPII: a membrane coat that forms endoplasmic reticulum‐derived vesicles , 1995, FEBS letters.

[40]  R. Klausner,et al.  Overexpression of wild-type and mutant ARF1 and ARF6: distinct perturbations of nonoverlapping membrane compartments , 1995, The Journal of cell biology.

[41]  S. Ferro-Novick,et al.  Ypt1p implicated in v-SNARE activation , 1994, Nature.

[42]  J. Rothman,et al.  A rab protein is required for the assembly of SNARE complexes in the docking of transport vesicles , 1994, Cell.

[43]  R. Schekman,et al.  COPII: A membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum , 1994, Cell.

[44]  J. Rothman,et al.  Implications of the SNARE hypothesis for intracellular membrane topology and dynamics , 1994, Current Biology.

[45]  R. Isberg,et al.  Two distinct defects in intracellular growth complemented by a single genetic locus in Legionella pneumophila , 1993, Molecular microbiology.

[46]  K. Joiner,et al.  Toxoplasma gondii: fusion competence of parasitophorous vacuoles in Fc receptor-transfected fibroblasts. , 1990, Science.

[47]  M. Zerial,et al.  Localization of low molecular weight GTP binding proteins to exocytic and endocytic compartments , 1990, Cell.

[48]  M. Muramatsu,et al.  A novel GTP-binding protein, Sar1p, is involved in transport from the endoplasmic reticulum to the Golgi apparatus , 1989, The Journal of cell biology.

[49]  P. Chardin,et al.  The human Rab genes encode a family of GTP-binding proteins related to yeast YPT1 and SEC4 products involved in secretion. , 1989, The Journal of biological chemistry.

[50]  G K Lewis,et al.  Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product , 1985, Molecular and cellular biology.

[51]  F. Maxfield,et al.  Legionella pneumophila inhibits acidification of its phagosome in human monocytes , 1984, The Journal of cell biology.

[52]  R. Schreiber,et al.  Evidence for a gamma-interferon receptor that regulates macrophage tumoricidal activity , 1984, The Journal of experimental medicine.

[53]  M. Horwitz.  The Legionnaires' disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes , 1983, The Journal of experimental medicine.

[54]  M. Horwitz.  Formation of a novel phagosome by the Legionnaires' disease bacterium (Legionella pneumophila) in human monocytes , 1983, The Journal of experimental medicine.

[55]  T. Puck,et al.  GENETICS OF SOMATIC MAMMALIAN CELLS , 1958, The Journal of experimental medicine.

[56]  T. Puck,et al.  GENETICS OF SOMATIC MAMMALIAN CELLS , 1956, The Journal of experimental medicine.

[57]  W. Balch,et al.  Partial Purification of the Sec 13 – 31 Complex from Rat Liver Cytosol , 1998 .