Identification of cellular proteins interacting with influenza A virus PB1-F2 protein.

UNLABELLED: The influenza A virus (IAV) protein PB1-F2, which is encoded by an alternative ORF of the PB1 polymerase gene, has been implicated as an important virulence factor and apoptosis inducer. However, the molecular mechanism of PB1-F2 function remains elusive. In this study, eight cellular proteins were identified as potential PB1-F2 interacting partners using the yeast two-hybrid system. Two positive candidate proteins, guanine nucleotide binding protein (G protein) beta polypeptide 2 (Gβ2) and macrophage migration inhibitory factor (MIF), were selected to be further characterized. The interaction of MIF and Gβ2 with PB1-F2 was confirmed by both GST pull-down and co-immunoprecipitation assays. Confocal laser microscopy showed that the interaction between PB1-F2 and the two cellular proteins occurred in the cytoplasm. The novel interactions between PB1-F2 and host proteins provide further pieces of evidence in the investigation of the pathogenic mechanism of IAV. KEYWORDS: influenza A virus; PB1-F2; yeast two-hybrid; protein-protein interaction.