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

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

Comments

Copyright 2018, Evan Daniel Kesinger. Used by permission

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