Quantitative Proteomic Analysis of A549 Cells Infected with Human Respiratory Syncytial Virus

Munday, Diane C.; Emmott, Edward; Surtees, Rebecca; Lardeau, Charles-Hugues; Wu, Weining; Duprex, W. Paul; Dove, Brian K.; Barr, John N.; Hiscox, Julian A.

Curiously, of a total of 510 proteins assigned to the nuclear proteome, 431 showed a 2-fold or greater decrease in abundance in HRSV-infected cells. This could not be attributed to a preparation or loading artifact as equal protein concentrations between the mock and infected preparations were validated using independent assays such as a BCA assay and staining after separation by one-dimensional SDS-PAGE, and large volumes were combined to reduce variation in liquid handling prior to LC-MS/MS analysis. Also, nuclear proteins with increased and decreased abundances between mock and infected cells were validated experimentally using alternative techniques on the same and different samples, e.g. the increased abundance of PML versus the decreased abundance of nucleolin and similar levels of marker proteins such as lamin B (e.g. Figs. 1 and 11) . A similar study of potential changes in the nuclear proteome in cells infected with an avian coronavirus reflected a general trend in the decreased abundance of nuclear proteins (37) .