Inhibition of influenza virus transcription by 2'-deoxy-2'-fluoroguanosine.
The nucleoside analog 2'-deoxy-2'-fluoroguanosine (2'-fluorodGuo) is phosphorylated by cellular enzymes and reversibly inhibits influenza virus replication in chick embryo cells within the first 4 h of infection. RNA hybridization studies revealed that primary and secondary transcription of influenza virus RNA were blocked at a compound concentration of 10 microM, but no inhibition of cell protein synthesis was seen even at high compound concentrations (200 microM). In vitro, the triphosphate of 2'-fluorodGuo is a competitive inhibitor of influenza virus transcriptase activity from disrupted virus, with a Ki of 1.0 microM. The cellular polymerases DNA polymerase alpha and RNA polymerase II were only weakly inhibited or were insusceptible to 2'-fluorodGTP. In kinetic studies with the influenza virus transcriptase, 2'-fluorodGTP, in the absence of GTP, blocked elongation of the virus RNA chain. Similarly, by using purified ribonucleoprotein complexes it was found that the addition of a single nucleotide of 2'-fluorodGTP to the virus RNA caused chain termination, which resulted in the blockage of further virus transcription. Furthermore, the specificity for influenza virus transcriptase was confirmed when the transcriptase from partially resistant virus was found to be 10-fold less susceptible to 2'-fluorodGTP (Ki = 13.1 microM).
Tisdale
M
M
Wellcome Research Laboratories, Beckenham, Kent, United Kingdom.
Ellis
M
M
Klumpp
K
K
Court
S
S
Ford
M
M
D000998:Antiviral Agents / Q000494:pharmacology*; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D009980:Influenza A virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D008715:Methionine / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D016865:Reassortant Viruses / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D012261:Ribonucleoproteins / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014529:Uridine / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D012333:RNA, Messenger; D012367:RNA, Viral; D012261:Ribonucleoproteins; D014764:Viral Proteins; C083670:2'-deoxy-2'-fluoroguanosine; D008715:Methionine; D012321:DNA-Directed RNA Polymerases; D003849:Deoxyguanosine; D014529:Uridine