Direct measurement of the influenza A virus M2 protein ion channel activity in mammalian cells.
The influenza A virus M2 integral membrane protein has an ion channel activity which is thought to play an essential role in the uncoating process of influenza virus in infected cells and, for some strains of influenza virus, in maintaining the hemagglutinin in its pH neutral form during transport through the trans Golgi network. To demonstrate directly that the M2 protein forms an ion channel in mammalian cells, the M2 protein was expressed in CV-1 cells by using an SV40-M2 recombinant virus and the whole cell membrane currents were recorded. It was found that the whole cell current was activated by low pH and inhibited by the M2 ion channel-specific blocker, amantadine hydrochloride. Expression of an altered M2 protein that contains a deletion of four residues in the transmembrane domain (M2-del28-31) and that when found in influenza virus confers amantadine resistance, resulted in a current that was activated by hyperpolarization of the membrane, was pH insensitive, and was resistant to block by amantadine. The data obtained in mammalian cells for the wild-type M2 and M2-del28-31 protein ion channel activities were very similar to those obtained when using the heterologous oocyte expression system.
Wang
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Howard Hughes Medical Institute, Northwestern University, Evanston, Illinois 60208.
Lamb
R A
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Pinto
L H
LH
D000547:Amantadine / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000378:metabolism*; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D008322:Mammals; D008564:Membrane Potentials / Q000235:genetics; D009154:Mutation; D014763:Viral Matrix Proteins / Q000235:genetics / Q000502:physiology*; D000086082:Viroporin Proteins
D000547:Amantadine; D007473:Ion Channels; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins