title: Analysis of A549 cell proteome alteration in response to recombinant influenza A virus nucleoprotein and its interaction with cellular proteins, a preliminary study.
abstract: Influenza A virus undergoes frequent changes of antigenicity and contributes to seasonal epidemics or unpredictable pandemics. Nucleoprotein, encoded by gene segment 5, is an internal protein of the virus and is conserved among strains of different host origins. In the current study, we analyzed the differentially expressed proteins in A549 cells transiently transfected with the recombinant nucleoprotein of influenza A virus by 2D gel electrophoresis. The resolved protein spots on gel were identified by MALDI-TOF/Mass spectrometry analysis. The majority of the host proteins detected to be differentially abundant in recombinant nucleoprotein-expressing cells as compared to vector-transfected cells are the proteins of metabolic pathways, glycolytic enzymes, molecular chaperones and cytoskeletal proteins. We further demonstrated the interaction of virus nucleoprotein with some of the identified host cellular proteins. In vitro binding assay carried out using the purified recombinant nucleoprotein (pET29a+NP-His) and A549 cell lysate confirmed the interaction between nucleoprotein and host proteins, such as alpha enolase 1, pyruvate kinase and β-actin. The preliminary data of our study provides the information on virus nucleoprotein interaction with proteins involved in glycolysis. However, studies are ongoing to understand the significance of these interactions in modulating the host factors during virus replication.
authors.forename: D
authors.initials: D
authors.lastname: Kumar
authors.forename: K
authors.initials: K
authors.lastname: Tiwari
authors.forename: M S
authors.initials: MS
authors.lastname: Rajala
mesh_terms: D000072283:A549 Cells; D000199:Actins / Q000235:genetics / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D009980:Influenza A virus / Q000378:metabolism*; D009698:Nucleoproteins / Q000235:genetics* / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D011770:Pyruvate Kinase / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; D009698:Nucleoproteins; D020543:Proteome; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; D011770:Pyruvate Kinase; C095157:ENO1 protein, human; D010751:Phosphopyruvate Hydratase
keywords: influenza A virus; A549 cells; nucleoprotein; alpha enolase 1; pyruvate kinase.