Activation of Gαi and subsequent uncoupling of receptor-Gαi signaling by Pasteurella multocida toxin

Joachim H C Orth, Ines Fester, Inga Preuss, Laura Agnoletto, Brenda A. Wilson, Klaus Aktories

Research output: Contribution to journalArticle

Abstract

Bacterial protein toxins are powerful tools for elucidating signaling mechanisms in eukaryotic cells. A number of bacterial protein toxins, e.g. cholera toxin, pertussis toxin (PTx), or Pasteurella multocida toxin (PMT), target heterotrimeric G proteins and have been used to stimulate or block specific signaling pathways or to demonstrate the contribution of their target proteins in cellular effects. PMT is a major virulence factor of P. multocida causing pasteurellosis in man and animals and is responsible for atrophic rhinitis in pigs. PMT modulates various signaling pathways, including phospholipase Cβ and RhoA, by acting on the heterotrimeric G proteins Gαq and Gα12/13, respectively. Here we report that PMT is a powerful activator of Gi protein. We show that PMT decreases basal isoproterenol and forskolin-stimulated cAMP accumulation in intact Swiss 3T3 cells, inhibits adenylyl cyclase activity in cell membrane preparations, and enhances the inhibition of cAMP accumulation caused by lysophosphatidic acid via endothelial differentiation gene receptors. PMT-mediated inhibition of cAMP production is independent of toxin activation of Gαq and/or Gα12/13. Although the effects of PMT are not inhibited by PTx, PMT blocks PTx-catalyzed ADP-ribosylation of Gi. PMT also inhibits steady-state GTPase activity and GTP binding of Gi in Swiss 3T3 cell membranes stimulated by lysophosphatidic acid. The data indicate that PMT is a novel activator of Gi, modulating its GTPase activity and converting it into a PTx-insensitive state.

Original languageEnglish (US)
Pages (from-to)23288-23294
Number of pages7
JournalJournal of Biological Chemistry
Volume283
Issue number34
DOIs
StatePublished - Aug 22 2008

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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