Detection of ion channel activity in Xenopus laevis oocytes expressing Influenza C virus CM2 protein

To demonstrate the ion channel activity of Influenza C virus CM2, we expressed this protein in oocytes of Xenopus laevis and measured whole cell currents by a two-electrode voltage-clamp method. It was found that the inward currents were induced upon hyperpolarizing the oocyte membranes. The amplitude of the currents increased slowly with time during the hyperpolarizing pulse, and the current-voltage relationship was nonlinear, having a slope that increased with the level of hyperpolarization. These results suggest strongly that CM2 forms a voltage-activated ion channel. The current amplitude was increased to a small extent by lowering the external pH. We also found that the anti-influenza A virus drug amantadine hydrochloride failed to attenuate the inward currents of CM2-expressing oocytes induced by hyperpolarization. The reversal voltage of tail currents was affected by the reduction of [Cl-], but neither by the change of [Na+] nor by that of [K+]. Furthermore, the amplitude of the inward currents was decreased by an anion channel blocker. The data presented here suggest that CM2 protein forms a voltage-activated ion channel permeable to chloride ion.