title: Myosin IIB deficiency in embryonic fibroblasts affects regulators and core members of the par polarity complex.
abstract: Wild-type (WT) and myosin heavy chain IIB null [MHCIIB (-/-)] embryonic fibroblasts were used as an experimental model to assess the role of the isoform B of myosin II (MII) in the regulation of the cell shape and intrinsic polarity. Genetic ablation of MHCIIB causes a persistent albeit, unstable protrusive activity in embryonic fibroblasts (Lo et al. in Nonmuscle myosin IIB is involved in the guidance of fibroblast migration. Mol Biol Cell 15:982-989, 2004). Here, we show that MHCIIB-deficient fibroblasts are characterized by a sustained guanine nucleotide exchange factor (GEF)-dependent activation of the small GTPase Rac-1 that is responsible for the continual lamellipodium formation. Moreover, we observed a sustained PKC-ζ activation and an increased association of cortactin with the plasma membrane in the MHCIIB (-/-) cells that were also dependent on GEF-mediated Rac-1 activation. Rac-1 activation and its downstream effects were induced in WT fibroblasts by inhibiting MII ATPase and crosslinking activities, suggesting that an altered actin-MII interaction favours Rac-1 activation, regardless of the MII isoform implicated. In addition, we found MIIB isoform-specific effects that were independent of Rac-1 activation. MHCIIA interacts with cortactin whereas MHCIIB does not. By contrast, MHCIIB interacts with Lgl1, a member of the Scribble/Dlg/Lgl polarity complex, whereas MHCIIA does not. MHCIIB (-/-) fibroblasts exhibited deregulated endogenous levels of the Par polarity complex members, Par3 and Par6. Together, the data show that MHCIIB deficiency causes imbalances in signalling pathways that are responsible for cell polarity determination. The results suggest that these pathways are targets of MIIB in the regulation of the cell's shape and polarity.
authors.affiliation: Department of Pathology and Cell Biology, Université de Montréal, Canada.
authors.forename: Sara
authors.initials: S
authors.lastname: Solinet
authors.forename: Casimir D
authors.initials: CD
authors.lastname: Akpovi
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Garcia
authors.forename: Ahmed
authors.initials: A
authors.lastname: Barry
authors.forename: María Leiza
authors.initials: ML
authors.lastname: Vitale
mesh_terms: D000818:Animals; D016764:Cell Polarity / Q000235:genetics*; D003600:Cytosol / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D020555:GTP-Binding Protein Regulators / Q000235:genetics / Q000378:metabolism; D020935:MAP Kinase Signaling System; D051379:Mice; D008822:Mice, Transgenic; D024501:Nonmuscle Myosin Type IIB / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D020555:GTP-Binding Protein Regulators; D024501:Nonmuscle Myosin Type IIB; D020830:rac1 GTP-Binding Protein