Histochemistry and Cell Biology

, 136:245 | Cite as

Myosin IIB deficiency in embryonic fibroblasts affects regulators and core members of the par polarity complex

  • Sara Solinet
  • Casimir D. Akpovi
  • Christopher J. Garcia
  • Ahmed Barry
  • María Leiza Vitale
Original Paper
First Online:
Accepted:

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.

Keywords

Myosin Actin Cortactin Polarity Rac1 
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Notes

Acknowledgments

We thankfully acknowledge the generous gift of the MHCIIB (−/−) and control mouse embryonic fibroblasts to Dr. R.S. Adelstein (Laboratory of Molecular Cardiology, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA). We thank Cytoskeleton Inc for the gift the G-LISA Cdc42 activation Assay Biochem Kit. We express our gratitude to Dr. R.-Marc Pelletier for his editorial assistance and helpful comments. We thank C. Charbonneau and M. Lauzon for their help in the confocal microscopy studies. This work was funded by the Natural Sciences and Engineering Research Council of Canada to MLV.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Sara Solinet
    • 1
  • Casimir D. Akpovi
    • 1
  • Christopher J. Garcia
    • 1
  • Ahmed Barry
    • 1
  • María Leiza Vitale
    • 1
  1. 1.Department of Pathology and Cell Biology, Faculty of MedicineUniversité de MontréalMontréalCanada

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