Investigation of the Downstream Regulation Mechanism of Beta-Interferon Using a High Coverage Quantitative Proteomic Workflow

Interferons have long been used for the treatment of a variety of cancers and viral infection diseases. However, we found in our previous study that a number of downsteam proteins of IFN were highly expressed in OSCC patients' tissue. Treating the cultured OSCC cell lines with IFN-β resulted in the enhanced cell mobility. Further quantitative proteomics and phosphoproteomics analyses revealed that in addition to the classical IFN-inducible proteins, a great number of proteins were also regulated by unknown mechanisms. Among those targets, several E3 ligases showed significant changes in either protein expression or phosphorylation levels. Such regulation mechanisms at degradation level have rare been discussed for IFN signaling pathways. Therefore, it is interesting to perform an in-depth analysis on this system. Since current proteomics approaches used in discovery phase possess low reproducibility, repeated experiments is suggested for getting reliable results. The common outcome is the reduction of the size of datasets. In this proposed project, we will establish a quantitative workflow which combines the shotgun approach with a directed strategy in the discovery phase. We expect that this strategy will increase the coverage of proteomes and reliability of quantification results when it is applied to the study of interferon triggered signaling events. Finally, the identified regulation mechanisms will be evaluated by using suitable inhibitors and antibody-based assays.

Project ID:PC10108-0289
External Project ID:NSC101-2320-B182-012