Mapping of functional domains on the influenza A virus RNA polymerase PB2 molecule using monoclonal antibodies.
Monoclonal antibodies against the PB2 of A/Puerto Rico/8/34 (A/PR/ 8/34) (H1N1) were prepared in order to define the functional domains of the RNA polymerase of influenza virus. The fifteen monoclonal antibodies that were generated were divided into 4 groups on the basis of ELISA binding to PB2 or its peptide fragments. Six Group I antibodies that bound to the PB2 N-terminal region (amino acids 1-104) did not inhibit transcription by the viral ribonucleoprotein complex. A single Group II antibody recognizing the region of amino acids 206-259 inhibited ApG-primed transcription. Groups III and IV antibodies bound to the C-terminal region of amino acids 660-759. Of these, Group III antibodies inhibited transcription. The present results identify multiple monoclonal antibody binding domains in PB2, two of which, when bound by antibodies, negatively affect viral RNA transcription.
Hatta
M
M
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Asano
Y
Y
Masunaga
K
K
Ito
T
T
Okazaki
K
K
Toyoda
T
T
Kawaoka
Y
Y
Ishihama
A
A
Kida
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001667:Binding, Competitive; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics; D018604:Epitope Mapping / Q000379:methods; D009980:Influenza A virus / Q000201:enzymology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012324:RNA-Dependent RNA Polymerase; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; C057027:PB2 protein, Influenzavirus A; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase; D012321:DNA-Directed RNA Polymerases