Production of the M2 protein of influenza A virus in insect cells is enhanced in the presence of amantadine

Abstract

Recombinant baculoviruses that express the M2 protein from the genes of either the amantadine-sensitive, influenza A/Ann Arbor/6/60 virus or a laboratoryderived, amantadine-resistant mutant of this virus were constructed. Addition of amantadine or rimantadine at 2 ~tg/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.

Functional reconstitution in lipid vesicles of influenza virus M2 protein expressed by baculovirus: evidence for proton transfer activity

“…Black et al (1993) reported a similar drug-induced enhancement of the expression of the M2 protein of influenza virus strain A/Ann Arbor/6/60, which differs from the Weybridge strain M2 by 15 amino acids (Ito et al, 1991). In contrast, amantadine treatment has no effect on M2 synthesis in virus-infected cells or M2 expression in mammalian cells or E. coli (F. Geraghty, A. Hayhurst & A. Hay, unpublished results).…”

Emergence of amantadine-resistant influenza A viruses: epidemiological study

“…[6][7][8] Emergence of amantadine-resistant strains Viral resistance to amantadine and its analogue rimantadine emerges quickly in vivo and in vitro. [1][2][3][4][5][9][10][11][12][13][14][15] Up to approximately one-third of patients may shed resistant viruses when amantadine or rimantadine is used for therapy. In vitro, sensitive viruses became resistant after three or five passages in the presence of 2 µg/ml amantadine.…”

Characterization of inhibition of M2 ion channel activity by BL-1743, an inhibitor of influenza A virus

“…Expression of the M 2 protein is deleterious to several host cell systems. Infection of Spodoptera frugiperda Sf9 cells with a recombinant baculovirus expressing the M 2 protein leads to inhibition of the replication of the recombinant baculovirus (1,26), and expression of M 2 protein in oocytes of X. laevis leads to premature cell death (5,13). In addition, expression of M 2 protein in Escherichia coli causes membrane permeability changes (6).…”