Interaction of potyvirus helper component-proteinase (HcPro) with RuBisCO and nucleosome in viral infections of plants.

Bean common mosaic virus (BCMV) causes severe disease in Phaseolus vulgaris plants. One of its non structural protein, the helper-component proteinase (HcPro) involves in multiple roles in aphid transmission, RNA binding, suppression of gene silencing and protease activity. The multifunctional role of HcPro hint towards its regulation at multiple host cellular sites. The mechanisms of these regulatory activities are poorly understood. Therefore, it is very important to study the molecular level interaction of HcPro with different cellular components. In this study, we demonstrate that the HcPro interacts with RuBisCo, an enzyme of chloroplast origin which might plays a crucial role in virus infection. A further line of experiments were carried out with factors of nuclear origin. Due to nucleic acid binding activity of HcPro, it showed interaction with dsDNA of nucleosome, as ascertained through electrophoretic mobility shift assay (EMSA). Interestingly, HcPro interacts with host nucleoprotein histones, H3 and H4. The gel-overlay assay and native electrophoresis-western blot analysis (NEWeB) revealed a direct interaction of BCMV HcPro with host nucleosome and with histones. These findings suggest that the BCMV through HcPro, not only utilize the host cytoplasmic components but also use host nuclear factors for its propagation and disease development.