Endosomal and secretory markers of the Legionella-containing vacuole.
The Gram-negative opportunistic pathogen Legionella pneumophila replicates in phagocytes within a specific compartment, the Legionella-containing vacuole (LCV). Formation of LCVs is a complex process requiring the bacterial Icm/Dot type IV secretion system and more than 100 translocated effector proteins, which putatively subvert cellular signaling and vesicle trafficking pathways. Phosphoinositide (PI) glycerolipids are pivotal regulators of signal transduction and membrane dynamics in eukaryotes. Recently, a number of Icm/Dot substrates were found to anchor to the LCV membrane by binding to PIs. One of these effectors, SidC, specifically interacts with phosphatidylinositol-4 phosphate [PtdIns(4)P]. Using an antibody against SidC and magnetic beads coupled to a secondary antibody, intact LCVs were purified by immuno-magnetic separation, followed by density centrifugation. This purification strategy is in principle applicable to any pathogen vacuole that carries specific markers. The LCV proteome determined by LC-MS/MS revealed 566 host proteins, including novel components of the endosomal pathway, as well as the early and late secretory trafficking pathways. Thus, LCV formation is a robust process that involves many (functionally redundant) Icm/Dot substrates, as well as the interaction with different host cell vesicle trafficking pathways.
Urwyler
Simon
S
Institute of Microbiology; ETH Zurich; Zurich, Switzerland.
Brombacher
Eva
E
Hilbi
Hubert
H
Traffic. 2005 Sep;6(9):711-9
16101675
Cell Microbiol. 2009 Feb;11(2):230-48
19016775
Environ Microbiol. 2007 Mar;9(3):563-75
17298357
Trends Microbiol. 2007 Aug;15(8):372-80
17632005
FEMS Microbiol Rev. 2005 Jan;29(1):65-81
15652976
Mol Biol Cell. 2002 Feb;13(2):656-69
11854420
Mol Microbiol. 2001 Nov;42(3):603-17
11722729
PLoS Pathog. 2006 Apr;2(4):e34
16652170
Infect Immun. 2005 Jul;73(7):4370-80
15972532
Cell Microbiol. 2009 Mar;11(3):442-60
19021631
Cell Microbiol. 2006 Nov;8(11):1697-706
16939534
Nat Cell Biol. 2006 Sep;8(9):971-7
16906144
Nat Rev Microbiol. 2009 Jan;7(1):13-24
19011659
Mol Microbiol. 2003 Apr;48(2):305-21
12675793
Mol Biol Cell. 2008 May;19(5):2059-68
18287531
Nature. 2007 Nov 15;450(7168):365-9
17952054
Traffic. 2007 May;8(5):618-28
17343681
Dev Cell. 2006 Nov;11(5):671-82
17084359
J Biol Chem. 2009 Feb 20;284(8):4846-56
19095644
Traffic. 2009 Jan;10(1):76-87
18980612
Cell Microbiol. 2008 Dec;10(12):2416-33
18673369
Dev Cell. 2006 Jul;11(1):47-56
16824952
Microbiology (Reading). 2004 Aug;150(Pt 8):2513-2525
15289548
PLoS Pathog. 2006 May;2(5):e46
16710455
Curr Opin Microbiol. 2006 Feb;9(1):86-94
16406773
Nat Cell Biol. 2002 Dec;4(12):945-54
12447391
Infect Immun. 2004 May;72(5):3048-53
15102819
Infect Immun. 2007 Feb;75(2):592-603
17101649
Science. 2002 Jan 25;295(5555):679-82
11809974
Cell Microbiol. 2008 Jun;10(6):1209-20
18363881
Science. 2007 Nov 9;318(5852):974-7
17947549
J Exp Med. 2004 May 3;199(9):1201-11
15117975
Mol Biol Cell. 1997 Jul;8(7):1343-60
9243512
Infect Immun. 2000 Sep;68(9):5154-66
10948139
Nat Cell Biol. 1999 Sep;1(5):280-7
10559940
Clin Microbiol Rev. 2002 Jul;15(3):506-26
12097254