The beta-dystroglycan (beta-DG) protein has the ability to target to multiple sites in eukaryotic cells, being a member of diverse protein assemblies including the transmembranal dystrophin-associated complex, and a nuclear envelope-localised complex that contains emerin and lamins A/C and B1. We noted that the importin alpha2/beta1-recognised nuclear localization signal (NLS) of beta-DG is also a binding site for the cytoskeletal-interacting protein ezrin, and set out to determine whether ezrin binding might modulate beta-DG nuclear translocation for the first time. Unexpectedly, we found that ezrin enhances rather than inhibits beta-DG nuclear translocation in C2C12 myoblasts. Both overexpression of a phosphomimetic activated ezrin variant (Ez-T567D) and activation of endogenous ezrin through stimulation of the Rho pathway resulted in both formation of actin-rich surface protrusions and significantly increased nuclear translocation of beta-DG as shown by quantitative microscopy and subcellular fractionation/Western analysis. In contrast, overexpression of a nonphosphorylatable inactive ezrin variant (Ez-T567A) or inhibition of Rho signaling, decreased nuclear translocation of beta-DG concomitant with a lack of cell surface protrusions. Further, a role for the actin cytoskeleton in ezrin enhancement of beta-DG nuclear translocation was implicated by the observation that an ezrin variant lacking its actin-binding domain failed to enhance nuclear translocation of beta-DG, while disruption of the actin cytoskeleton led to a reduction in beta-DG nuclear localization. Finally, we show that ezrin-mediated cytoskeletal reorganization enhances nuclear translocation of the cytoplasmic but not the transmembranal fraction of beta-DG. This is the first study showing that cytoskeleton reorganization can modulate nuclear translocation of beta-DG, with the implication that beta-DG can respond to cytoskeleton-driven changes in cell morphology by translocating from the cytoplasm to the nucleus to orchestrate nuclear processes in response to the functional requirements of the cell.
Vasquez-Limeta, A., Wagstaff, K. M., Ortega, A., Crouch, D. H., Jans, D. A., & Cisneros, B. (2014). Nuclear import of beta-dystroglycan is facilitated by ezrin-mediated cytoskeleton reorganization. PLoS ONE, 9(3 (Art. No: e90629)), 1 - 13. https://doi.org/10.1371/journal.pone.0090629