title: Pasteurella multocida toxin prevents osteoblast differentiation by transactivation of the MAP-kinase cascade via the Gα(q/11)--p63RhoGEF--RhoA axis.
abstract: The 146-kDa Pasteurella multocida toxin (PMT) is the main virulence factor to induce P. multocida-associated progressive atrophic rhinitis in various animals. PMT leads to a destruction of nasal turbinate bones implicating an effect of the toxin on osteoblasts and/or osteoclasts. The toxin induces constitutive activation of Gα proteins of the G(q/11)-, G₁₂/₁₃- and G(i)-family by deamidating an essential glutamine residue. To study the PMT effect on bone cells, we used primary osteoblasts derived from rat calvariae and stromal ST-2 cells as differentiation model. As marker of functional osteoblasts the expression and activity of alkaline phosphatase, formation of mineralization nodules or expression of specific transcription factors as osterix was determined. Here, we show that the toxin inhibits differentiation and/or function of osteoblasts by activation of Gα(q/11). Subsequently, Gα(q/11) activates RhoA via p63RhoGEF, which specifically interacts with Gα(q/11) but not with other G proteins like Gα₁₂/₁₃ and Gα(i). Activated RhoA transactivates the mitogen-activated protein (MAP) kinase cascade via Rho kinase, involving Ras, MEK and ERK, resulting in inhibition of osteoblast differentiation. PMT-induced inhibition of differentiation was selective for the osteoblast lineage as adipocyte-like differentiation of ST-2 cells was not hampered. The present work provides novel insights, how the bacterial toxin PMT can control osteoblastic development by activating heterotrimeric G proteins of the Gα(q/11)-family and is a molecular pathogenetic basis for understanding the role of the toxin in bone loss during progressive atrophic rhinitis induced by Pasteurella multocida.
authors.affiliation: Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Albert-Ludwigs-Universität Freiburg, Freiburg, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Siegert
authors.forename: Gudula
authors.initials: G
authors.lastname: Schmidt
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Papatheodorou
authors.forename: Thomas
authors.initials: T
authors.lastname: Wieland
authors.forename: Klaus
authors.initials: K
authors.lastname: Aktories
authors.forename: Joachim H C
authors.initials: JH
authors.lastname: Orth
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D051379:Mice; D010006:Osteoblasts / Q000378:metabolism* / Q000473:pathology; D010014:Osteolysis / Q000378:metabolism / Q000473:pathology; D010326:Pasteurella Infections / Q000378:metabolism* / Q000473:pathology; D016979:Pasteurella multocida / Q000378:metabolism* / Q000472:pathogenicity; D051381:Rats; D012222:Rhinitis, Atrophic / Q000378:metabolism / Q000473:pathology; D064067:Rho Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D012886:Skull / Q000378:metabolism / Q000473:pathology; D017154:Stromal Cells / Q000378:metabolism / Q000473:pathology; D015533:Transcriptional Activation; D037521:Virulence Factors / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; C577427:GEFT protein, mouse; C062258:Pasteurella multocida toxin; D064067:Rho Guanine Nucleotide Exchange Factors; D037521:Virulence Factors; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; C526380:RhoA protein, mouse; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein