cAMP regulates Ca2+-dependent exocytosis of lysosomes and lysosome-mediated cell invasion by trypanosomes.
Ca2+-regulated exocytosis, previously believed to be restricted to specialized cells, was recently recognized as a ubiquitous process. In mammalian fibroblasts and epithelial cells, exocytic vesicles mobilized by Ca2+ were identified as lysosomes. Here we show that elevation in intracellular cAMP potentiates Ca2+-dependent exocytosis of lysosomes in normal rat kidney fibroblasts. The process can be modulated by the heterotrimeric G proteins Gs and Gi, consistent with activation or inhibition of adenylyl cyclase. Normal rat kidney cell stimulation with isoproterenol, a beta-adrenergic agonist that activates adenylyl cyclase, enhances Ca2+-dependent lysosome exocytosis and cell invasion by Trypanosoma cruzi, a process that involves parasite-induced [Ca2+]i transients and fusion of host cell lysosomes with the plasma membrane. Similarly to what is observed for T. cruzi invasion, the actin cytoskeleton acts as a barrier for Ca2+-induced lysosomal exocytosis. In addition, infective stages of T. cruzi trigger elevation in host cell cAMP levels, whereas no effect is observed with noninfective forms of the parasite. These findings demonstrate that cAMP regulates lysosomal exocytosis triggered by Ca2+ and a parasite/host cell interaction known to involve Ca2+-dependent lysosomal fusion.
Rodríguez
A
A
Departments of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, 06520, USA.
Martinez
I
I
Chung
A
A
Berlot
C H
CH
Andrews
N W
NW
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000199:Actins / Q000502:physiology; D000067956:Adenylyl Cyclase Inhibitors; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology*; D003599:Cytoskeleton / Q000502:physiology; D003600:Cytosol / Q000378:metabolism; D005089:Exocytosis / Q000187:drug effects*; D005347:Fibroblasts / Q000166:cytology; D019204:GTP-Binding Proteins / Q000378:metabolism; D007097:Imines / Q000494:pharmacology; D007545:Isoproterenol / Q000494:pharmacology; D007668:Kidney / Q000166:cytology; D008247:Lysosomes / Q000378:metabolism*; D008561:Membrane Fusion; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D051381:Rats; D014349:Trypanosoma cruzi / Q000472:pathogenicity*
D000199:Actins; D000067956:Adenylyl Cyclase Inhibitors; D000318:Adrenergic beta-Agonists; D007097:Imines; D010726:Phosphodiesterase Inhibitors; C014925:RMI 12330A; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D007545:Isoproterenol; D002118:Calcium; D015056:1-Methyl-3-isobutylxanthine