<i>Pasteurella Multocida</i> Toxin Prevents Osteoblast Differentiation by Transactivation of the MAP-Kinase Cascade via the Gα_q/11 - p63RhoGEF - RhoA Axis

<div><p>The 146-kDa <i>Pasteurella multocida</i> toxin (PMT) is the main virulence factor to induce <i>P. multocida</i>-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-, G_12/13- 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 Gα_12/13 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 <i>Pasteurella multocida</i>.</p></div