Department of Plant Pathology, Washington State University, Pullman, WA, USA.
Sanya Institute of Breeding and Multiplication/School of Tropical Agriculture and Forestry, Hainan University, Sanya, China.
Int J Biol Macromol. 2024 Jun;269(Pt 2):132095. doi: 10.1016/j.ijbiomac.2024.132095. Epub 2024 May 4.
Plant viruses are the most abundant destructive agents that exist in every ecosystem, causing severe diseases in multiple crops worldwide. Currently, a major gap is present in computational biology determining plant viruses interaction with its host. We lay out a strategy to extract virus-host protein interactions using various protein binding and interface methods for Geminiviridae, a second largest virus family. Using this approach, transcriptional activator protein (TrAP/C2) encoded by Cotton leaf curl Kokhran virus (CLCuKoV) and Cotton leaf curl Multan virus (CLCuMV) showed strong binding affinity with calmodulin-like (CML) protein of Gossypium hirsutum (Gh-CML11). Higher negative value for the change in Gibbs free energy between TrAP and Gh-CML11 indicated strong binding affinity. Consensus from gene ontology database and in-silico nuclear localization signal (NLS) tools identified subcellular localization of TrAP in the nucleus associated with Gh-CML11 for virus infection. Data based on interaction prediction and docking methods present evidences that full length and truncated C2 strongly binds with Gh-CML11. This computational data was further validated with molecular results collected from yeast two-hybrid, bimolecular fluorescence complementation system and pull down assay. In this work, we also show the outcomes of full length and truncated TrAP on plant machinery. This is a first extensive report to delineate a role of CML protein from cotton with begomoviruses encoded transcription activator protein.
植物病毒是存在于每个生态系统中最丰富的破坏性因素，导致全球多种作物发生严重疾病。目前，在计算生物学领域存在一个主要的差距，即确定植物病毒与其宿主相互作用的机制。我们提出了一种使用各种蛋白质结合和界面方法来提取双生病毒科（第二大病毒家族）病毒-宿主蛋白相互作用的策略。使用这种方法，棉花曲叶病毒（CLCuKoV）和棉花曲叶多利病毒（CLCuMV）编码的转录激活蛋白（TrAP/C2）与棉属Gh-CML11 的钙调素样蛋白（CML）显示出很强的结合亲和力。TrAP 与 Gh-CML11 之间吉布斯自由能变化的负值较高，表明结合亲和力较强。来自基因本体数据库和计算机核定位信号（NLS）工具的共识确定了 TrAP 在细胞核中的亚细胞定位，与 Gh-CML11 相关，以利于病毒感染。基于相互作用预测和对接方法的数据提供了证据，全长和截短的 C2 与 Gh-CML11 强烈结合。这些计算数据通过酵母双杂交、双分子荧光互补系统和下拉测定收集的分子结果得到进一步验证。在这项工作中，我们还展示了全长和截短的 TrAP 对植物机制的影响。这是首次详细报告了棉属 CML 蛋白与双生病毒编码转录激活蛋白的作用。
