Production of the M2 protein of influenza A virus in insect cells is enhanced in the presence of amantadine.
Recombinant baculoviruses that express the M2 protein from the genes of either the amantadine-sensitive, influenza A/Ann Arbor/6/60 virus or a laboratory-derived, amantadine-resistant mutant of this virus were constructed. Addition of amantadine or rimantadine at 2 micrograms/ml to cultures of Sf9 cells infected with the recombinant baculoviruses increased the yield of the M2 protein from the amantadine-sensitive virus approximately 10-fold, but did not increase the yield of the M2 protein from the amantadine-resistant virus. Flow cytometry demonstrated that the increased production of M2 in the presence of amantadine resulted in increased cell surface expression of the M2 protein. Pulse-chase experiments indicated that whereas the rate of synthesis of the M2 protein increased in the presence of amantadine, the M2 protein was stable in both the presence and absence of amantadine. Addition of amantadine to Sf9 cells as late as 72 h after infection with the recombinant virus increased the production of M2 protein. These data suggest that the M2 protein exerts some biological activity in Sf9 cells.
Black
R A
RA
Influenza Branch, Centers for Disease Control, Atlanta, Georgia 30333.
Rota
P A
PA
Gorodkova
N
N
Cramer
A
A
Klenk
H D
HD
Kendal
A P
AP
D000547:Amantadine / Q000494:pharmacology*; D000818:Animals; D016367:Baculoviridae; D002478:Cells, Cultured; D015870:Gene Expression / Q000502:physiology; D009980:Influenza A virus / Q000187:drug effects*; D009036:Moths; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000187:drug effects; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000187:drug effects*; D000086082:Viroporin Proteins
D000547:Amantadine; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins