Mobilization of protein kinase C in macrophages induced by Listeria monocytogenes affects its internalization and escape from the phagosome.
Listeriolysin O (LLO) and a phosphatidylinositol-specific phospholipase C (PI-PLC) are known virulence factors of Listeria monocytogenes in both tissue cultures and the murine model of infection. LLO is a member of a family of pore-forming cholesterol-dependent cytotoxins and is known to play an essential role in escape from the primary phagocytic vacuole of macrophages. PI-PLC plays an accessory role, in that PI-PLC mutants are partially defective in escape. We have shown that both of these molecules are essential for initiating rapid increases in the calcium level in the J774 murine macrophage cell line (S. J. Wadsworth and H. Goldfine, Infect. Immun. 67:1770-1778, 1999). Here we show that both LLO and PI-PLC are required for translocation of protein kinase C delta (PKC delta) to the periphery of J774 cells and for translocation of PKC beta II to early endosomes beginning within the first minute after addition of bacteria to the culture medium. Treatment with the calcium channel blocker SK&F 96365 inhibited translocation of PKC beta II but not PKC delta. Our findings lead us to propose a host signaling pathway requiring LLO and the formation of diacylglycerol by PI-PLC in which calcium-independent PKC delta is responsible for the initial calcium signal and the subsequent PKC beta II translocation. LLO-dependent translocation of PKC beta I to early endosomes also occurs between 1 and 4 min after infection, but this occurs in the absence of PI-PLC. All of these signals were observed in cells that had not internalized bacteria. Blocking PKC beta translocation with hispidin resulted in more rapid uptake of wild-type bacteria and greatly reduced escape from the primary phagocytic vacuoles of J774 cells.
Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia 19104-6076, USA.
Sandra J
SJ
Wadsworth
Howard
H
Goldfine
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology; D001427:Bacterial Toxins; D001578:Benzopyrans / Q000494:pharmacology; D001692:Biological Transport; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002460:Cell Line; D011992:Endosomes / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006360:Heat-Shock Proteins / Q000235:genetics / Q000276:immunology; D006460:Hemolysin Proteins; D007093:Imidazoles / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000276:immunology*; D008089:Listeria monocytogenes / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008264:Macrophages / Q000166:cytology / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D010587:Phagocytosis / Q000276:immunology*; D010588:Phagosomes / Q000276:immunology / Q000382:microbiology*; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000276:immunology*; D064546:Protein Kinase C beta; D051745:Protein Kinase C-delta; D011753:Pyrones / Q000494:pharmacology; D010738:Type C Phospholipases / Q000235:genetics / Q000276:immunology; D014617:Vacuoles
D000098:Acetophenones; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D001578:Benzopyrans; D002121:Calcium Channel Blockers; D004791:Enzyme Inhibitors; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D007093:Imidazoles; D007527:Isoenzymes; D011753:Pyrones; C085746:rottlerin; C497259:Prkcd protein, mouse; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051745:Protein Kinase C-delta; D010738:Type C Phospholipases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase; C063159:1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole; C038705:hlyA protein, Listeria monocytogenes; C007946:hispidin