Influenza virus PB1 protein is the minimal and essential subunit of RNA polymerase

RNA polymerase of influenza virus with the subunit structure PB1-PB2-PA is involved in both transcription and replication of the genome RNA. The RNA polymerase with transcription activity was reconstituted from three P proteins, which were separately isolated from insect cells infected with recombinant baculoviruses, each carrying cDNA for one P protein. Nuclear extracts of the insect cells infected with each of the recombinant baculoviruses or various combinations of these viruses were examined for transcription and replication activities. The nuclear extract of cells expressing all three P proteins catalyzed model template-directed RNA synthesis in the absence of primers (an indication of RNA replication), supporting the notion that the complete set of three P proteins is required for RNA replication. All the nuclear extracts containing the PB1 subunit, including the extract containing PB1 alone, were able to catalyze model template-directed dinucleotide-primed RNA synthesis (an indication of transcription). These observations not only confirm that the PB1 protein is a catalytic subunit of influenza virus RNA polymerase, but also indicate that PB1 alone is able to catalyze RNA synthesis in the absence of PB2 and PA subunits.