Influenza D Virus M2 Protein Exhibits Ion Channel Activity in Xenopus laevis Oocytes

Authors

• Evan Daniel Kesinger, University of Nebraska-LincolnFollow

First Advisor

Hideaki Moriyama

Date of this Version

4-2018

Document Type

Thesis

Citation

Kesinger, Evan (2018) Influenza D Virus M2 Protein Exhibits Ion Channel Activity in Xenopus laevis Oocytes. dissertations and Theses in Biological Sciences.

Comments

A thesis Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science Major: Biological Sciences Under the Supervision of Professor Hideaki Moriyama Lincoln, Nebraska April, 2018.

Copyright (c) 2018 Evan Daniel Kesinger

Abstract

The Influenza virus M2 ion channel has good potential as a target for antiviral drugs, as the channel is necessary for viral replication. M2 of Influenza A and B viruses has beenstudied extensively, and is understood to function as a proton channel. Antiviral drugs like amantadine and rimantadine have been used to block the function of Influenza A virus M2. Influenza C virus M2 has also been researched and is understood to act as a chloride ion channel. However, the M2 channel of Influenza D virus (DM2) has been studied very little, and the activity and mechanism of the channel are unknown. To test the function of the channel, the protein was expressed in a Xenopus laevis oocyte system, and expression was confirmed with immunofluorescence and mass spectrometry. DM2 expressed in oocytes was analyzed using a two-electrode voltage clamp. The native DM2 protein was tested along with C-terminal altered M2 and M2 altered at an intracellular motif. The native DM2 channel was found to exhibit ion channel activity and chloride specificity comparable to that of Influenza C virus M2, and C-terminal and transmembrane altered M2 was found to exhibit altered ion channel activity from native DM2. These results suggest that DM2 functions as a chloride ion channel.

Advisor: Hideaki Moriyama