Characteristics of arbidol-resistant mutants of influenza virus: implications for the mechanism of anti-influenza action of arbidol.
The antiviral drug arbidol (ARB), which is licensed in Russia for use against influenza, is known to inhibit early membrane fusion events in influenza A and B virus replication. To investigate in more detail the target and mechanism of ARB action we generated and studied the characteristics of ARB-resistant influenza virus mutants. Observations of the ARB susceptibility of reassortants between A/Singapore/1/57(H2N2) and A/chicken/Germany/27(H7N7, "Weybridge" strain) and of mutants of the latter virus identified the virus haemagglutinin (HA) as the major determinant of ARB sensitivity. ARB-resistant mutants, selected from the most sensitive reassortant, possessed single amino acid substitutions in the HA2 subunit which caused an increase in the pH of fusion and the associated conformational change in HA. ARB was shown to stabilize the HA by causing a 0.2 pH unit reduction in the pH of the transition to the low pH form, which was specifically abrogated by the resistance mutations. Some of the resistance mutations, which reduce acid stability and would disrupt ARB-HA interactions, are located in the vicinity of a potential ARB binding site identified using the docking programme Gold. Together, the results of these investigations indicate that ARB falls within a class of inhibitor which interacts with HA to stabilize it against the low pH transition to its fusogenic state and consequently inhibit HA-mediated membrane fusion during influenza virus infection.
Leneva
Irina A
IA
National Institute for Medical Research, Mill Hill, London NW7 1AA, UK. wnyfd385@yandex.ru
Russell
Rupert J
RJ
Boriskin
Yury S
YS
Hay
Alan J
AJ
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D004285:Dogs; D024882:Drug Resistance, Viral; D006389:Hemagglutinins, Viral / Q000737:chemistry / Q000235:genetics; D007211:Indoles / Q000494:pharmacology*; D053121:Influenza A Virus, H2N2 Subtype / Q000187:drug effects*; D053126:Influenza A Virus, H7N7 Subtype / Q000187:drug effects*; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D020125:Mutation, Missense; D011487:Protein Conformation; D016865:Reassortant Viruses / Q000187:drug effects*; D013440:Sulfides
D000998:Antiviral Agents; D006389:Hemagglutinins, Viral; D007211:Indoles; C452559:hemagglutinin HA-2 fusogenic peptide, Influenza virus; C086979:umifenovir; D013440:Sulfides