Effects of large clostridial cytotoxins on activation of RBL 2H3-hm1 mast cells indicate common and different roles of Rac in FcepsilonRI and M1-receptor signaling.
Using Rho GTPases-inhibiting clostridial cytotoxins, we showed recently in RBL cells that the GTPase Rac is involved in FcepsilonRI (high-affinity receptor for IgE) signaling and receptor-mediated calcium mobilization, including influx via calcium release-activated calcium channels. Here, we studied the role of Rho GTPases in muscarinic M1 receptor signaling in RBL 2H3-hm1 cells. Clostridium difficile toxin B, which inactivates Rho, Rac, and Cdc42, and Clostridium sordellii lethal toxin, which inhibits Rac but not Rho, blocked M1-mediated exocytosis, indicating that Rac but not Rho is involved in the regulation of receptor-mediated exocytosis. Although antigen-induced FcepsilonRI stimulation caused tyrosine phosphorylation of the Rac guanine nucleotide exchange factor Vav, M1 stimulation by carbachol activated Rac independently of Vav. The Rac-inactivating toxins blocked M1 receptor-induced membrane translocation of the pleckstrin homology domain of protein kinase B, which is a phosphoinositide 3-kinase effector. The M1-induced calcium release from internal stores was not affected by toxin B; however, the subsequent calcium influx from the extracellular space was inhibited. The data suggest that besides capacitative calcium entry, the M1 signaling pathway activates further calcium entry channels with mechanisms that are not affected by the inhibition of Rac.
Djouder
Nabil
N
Institut für Experimentelle und Klinische Pharmakologie, Universität Freiburg, Freiburg, Germany.
Aneiros
Eduardo
E
Cavalié
Adolfo
A
Aktories
Klaus
K
D000818:Animals; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D006801:Humans; D008407:Mast Cells; D051379:Mice; D043643:Receptor, Muscarinic M1; D017455:Receptors, IgE; D011976:Receptors, Muscarinic; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection; D014407:Tumor Cells, Cultured; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism; D020744:rac GTP-Binding Proteins
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D020662:Guanine Nucleotide Exchange Factors; D043643:Receptor, Muscarinic M1; D017455:Receptors, IgE; D011976:Receptors, Muscarinic; C031815:toxB protein, Clostridium difficile; D001619:beta-N-Acetylhexosaminidases; D020744:rac GTP-Binding Proteins