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

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Interaction of potyvirus helper component-proteinase (HcPro) with RuBisCO and nucleosome in viral infections of plants.
Plant Physiology and Biochemistry ( IF 6.1 ) Pub Date : 2020-03-31 , DOI: 10.1016/j.plaphy.2020.03.036
Sunil Kumar ₁ , Ruma Karmakar ₂ , Ishu Gupta ₁ , Ashok Kumar Patel ₁

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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.

中文翻译：

马铃薯Y病毒辅助成分蛋白酶（HcPro）与RuBisCO和核小体在植物病毒感染中的相互作用。

菜豆普通花叶病毒 (BCMV) 会导致菜豆植物严重患病。辅助成分蛋白酶 (HcPro) 是其非结构蛋白之一，在蚜虫传播、RNA 结合、基因沉默抑制和蛋白酶活性中发挥多种作用。 HcPro 的多功能作用暗示了它在多个宿主细胞位点的调节。人们对这些监管活动的机制知之甚少。因此，研究HcPro与不同细胞成分的分子水平相互作用非常重要。在这项研究中，我们证明 HcPro 与 RuBisCo 相互作用，RuBisCo 是一种叶绿体来源的酶，可能在病毒感染中发挥至关重要的作用。利用核源因子进行了进一步的实验。通过电泳迁移率变动分析 (EMSA) 确定，由于 HcPro 的核酸结合活性，它显示出与核小体 dsDNA 的相互作用。有趣的是，HcPro 与宿主核蛋白组蛋白 H3 和 H4 相互作用。凝胶覆盖测定和天然电泳-蛋白质印迹分析 (NEWeB) 揭示了 BCMV HcPro 与宿主核小体和组蛋白的直接相互作用。这些发现表明，BCMV 通过 HcPro 不仅利用宿主细胞质成分，还利用宿主核因子进行繁殖和疾病发展。

更新日期：2020-03-31

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