Current Approaches on Viral Infection: Proteomics and Functional Validations

Zheng, Jie; Tan, Boon Huan; Sugrue, Richard; Tang, Kai

In addition, quantitative proteomics could reveal virus-host interactions with distinctive profiles after plasmid transfections. The X protein of chronic hepatitis B virus (HBx) was known to induce hepatocellular carcinoma (HCC). In one study, HBx of genotype A, B, C were amplified by PCR and inserted into pXJ40 vector, followed by transfection HepG2 cells (Feng et al., 2010) . Cells transfected with those different genotypes of HBx and empty pXJ40 plasmid were labeled with iTRAQ and further submitted to 2D LC-MS/MS. Comprehensive protein profiling displayed some up-regulated cytoskeleton proteins responsible for cytoskeleton movement and migration. For instance, microtubule-actin crosslinking factor 1 (MACF1), annexin A2, high mobility group box 1 (HMGB1), were over-expressed after HBx transfection. To further functionally validate the proteomic data in respect to cellular motility, the HepG2 cells were co-transfected by those HBx genotypes and green fluorescent protein (GFP) to visualize the tracks of cell movement by real-time fluorescence microscopy. And HBx genotype A infected cells revealed more vigorous movement than cells infected with other HBx genotypes. Similarly, Mota et al.