Viral proteomics: The emerging cutting-edge of virus research

Zhou, ShengTao; Liu, Rui; Zhao, Xia; Huang, CanHua; Wei, YuQuan

Although the Y2H system provides a high-throughput tool for protein-protein interaction screen, it cannot monitor the dynamic interactions that occur upon virus infection. One group constructed a modified immunoprecipitation protocol to unravel the dynamic viral protein interaction using a green fluorescent protein (GFP)-tagged virus [44] . This protocol involved tagging the virus with GFP-encoded proteins, visualizing the proteins' locations in the host cell at distinct infection stages, isolating interacting proteins using GFP affinity columns, and protein validation by MS. This study examined the interaction of the nsP3 protein of αvirus, Sindbis, with mammalian proteins at different times during infection. They demonstrated that a host factor was recruited to nsP3-containing complexes in a time-dependent fashion. This process was concomitant with a specific early and continuous recruitment of G3BP, a nuclear transport factor, and a subsequent recruitment of 14-3-3 proteins. These conclusions not only confirmed that Sindbis virus infection can modify RNA transport, but more significantly, revealed the utility of novel high-throughput methodologies to verify protein-protein interactions in host cells infected by viruses.