Dynamin 2 interacts with connexin 26 to regulate its degradation and function in gap junction formation

•We identified dynamin 2 (Dyn2) as a Cx26 interactor in yeast and mammalian cells.•Interaction involves the C-terminus of Cx26 and the GTPase effector domain of Dyn2.•Dyn2 inhibition using siRNA or dynasore resulted in reduced Cx26 degradation.•Dyn2 regulates Cx26 endocytosis and ubiquitination.•Our results establish Dyn2 as a Cx26 partner in the regulation of GJIC. Connexin 26 (Cx26), a protein involved in gap junctional intercellular communication, has an essential function during organ and tissue development. Its deregulation, in part due to inherent mutations, is associated with pathological conditions including congenital deafness. Regulation of Cx26 protein level is critical for its function but the molecular mechanisms involved are partially understood. This study identifies dynamin 2 (Dyn2) as a Cx26 interactor in yeast and mammalian cells. Deletion studies revealed that Cx26–Dyn2 interaction involves the C-terminus of Cx26 and the GTPase effector domain of Dyn2, which is of particular importance for the regulation of the endocytic pathway. Dyn2 inhibition using siRNA or dynasore resulted in reduced Cx26 degradation at the plasma membrane and this was associated with change in gap junctional intercellular communication (GJIC). Furthermore, we demonstrate that Dyn2 regulates Cx26 endocytosis and ubiquitination. These results establish Dyn2 as a Cx26 partner in the regulation of GJIC.