Abstract
Three recombinant influenza A viruses with different neuraminidases (NAs) in the background of A/PR/8/34 (PR8), named rPR8-H5N1NA, rPR8-H9N2NA, and rPR8-H1N1NA, derived from H5N1, H9N2, H1N1 (swine) viruses, respectively, were constructed. We performed a quantitative proteomics analysis to investigate differential protein expression in Madin-Darby canine kidney (MDCK) cells infected with recombinant and wild-type influenza viruses to determine whether NA replacement would alter host cell gene expression. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-TOF MS) and two-dimensional gel electrophoresis (2-DE), we identified 12 up-regulated and 49 down-regulated protein spots, including cytoskeletal proteins, molecular biosynthesis proteins, ubiquitin-proteasome pathway proteins, and heat shock proteins. The most significant changes in infected cells were observed for molecular biosynthesis proteins. We found more differentially expressed protein spots in cells infected with rPR8-H5N1NA or rPR8-H9N2NA viruses than cells infected with wild-type virus. Many of those proteins are postulated to be involved in cell-cell fusion, but the full mechanism remains to be explored. Meanwhile, our data demonstrate that the wild-type virus has evolutionary advantages over recombinant viruses.
Highlights
Virus evolution is inseparable from virus–host interactions, and there have been many studies focused on the interactions between influenza viruses and their hosts in the past several years [1]
We found that 61 proteins in the infected cells differed between cells infected with wild-type virus versus those infected with recombinant viruses, including 12 significantly up-regulated proteins and 49 significantly down-regulated proteins (Fig. 1C)
More differential spots were identified in cells infected with rPR8-H5N1NA or rPR8H9N2NA recombinant viruses than those infected with other viruses (Fig. 1B)
Summary
Virus evolution is inseparable from virus–host interactions, and there have been many studies focused on the interactions between influenza viruses and their hosts in the past several years [1]. It is unknown whether proteins will be differentially expressed in MDCK cells infected with recombinant viruses versus wild-type virus and which proteins will be affected. Statistical analysis The relative spot intensities corresponding to the levels of abundance of proteins in the influenza virus-infected and mockinfected groups or recombinant viruses-infected and PR8-wtinfected groups were compared.
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