Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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GTP-Binding Proteins / genetics
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GTP-Binding Proteins / metabolism*
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Humans
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Influenza A Virus, H1N1 Subtype / genetics
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Influenza A Virus, H1N1 Subtype / metabolism*
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Influenza, Human / genetics
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Influenza, Human / metabolism*
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Influenza, Human / virology
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Intramolecular Oxidoreductases / genetics
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Intramolecular Oxidoreductases / metabolism*
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Macrophage Migration-Inhibitory Factors / genetics
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Macrophage Migration-Inhibitory Factors / metabolism*
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Neoplasm Proteins / genetics
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Neoplasm Proteins / metabolism*
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Protein Binding
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Receptors for Activated C Kinase
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Receptors, Cell Surface / genetics
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Receptors, Cell Surface / metabolism*
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Two-Hybrid System Techniques
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Viral Proteins / genetics
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Viral Proteins / metabolism*
Substances
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GNB2 protein, human
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Macrophage Migration-Inhibitory Factors
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Neoplasm Proteins
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PB1-F2 protein, Influenza A virus
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RACK1 protein, human
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Receptors for Activated C Kinase
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Receptors, Cell Surface
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Viral Proteins
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GTP-Binding Proteins
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Intramolecular Oxidoreductases
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MIF protein, human