Centro de Biotecnología y Genómica de Plantas (UPM-INIA), Pozuelo de Alarcón, Spain.
Instituto de Biología Molecular y Celular de Plantas (UPV-CSIC), IBMCP, Valencia, Spain.
Mol Plant Pathol. 2020 Oct;21(10):1271-1286. doi: 10.1111/mpp.12976. Epub 2020 Aug 1.
Virus infections affect plant developmental traits but this aspect of the interaction has not been extensively studied so far. Two strains of Turnip mosaic virus differentially affect Arabidopsis development, especially flower stalk elongation, which allowed phenotypical, cellular, and molecular characterization of the viral determinant, the P3 protein. Transiently expressed wild-type green fluorescent protein-tagged P3 proteins of both strains and selected mutants of them revealed important differences in their behaviour as endoplasmic reticulum (ER)-associated peripheral proteins flowing along the reticulum, forming punctate accumulations. Three-dimensional (3D) model structures of all expressed P3 proteins were computationally constructed through I-TASSER protein structure predictions, which were used to compute protein surfaces and map electrostatic potentials to characterize the effect of amino acid changes on features related to protein interactions and to phenotypical and subcellular results. The amino acid at position 279 was the main determinant affecting stalk development. It also determined the speed of ER-flow of the expressed proteins and their final location. A marked change in the protein surface electrostatic potential correlated with changes in subcellular location. One single amino acid in the P3 viral protein determines all the analysed differential characteristics between strains differentially affecting flower stalk development. A model proposing a role of the protein in the intracellular movement of the viral replication complex, in association with the viral 6K2 protein, is proposed. The type of association between both viral proteins could differ between the strains.
病毒感染会影响植物的发育特征,但到目前为止,这种相互作用的方面还没有得到广泛研究。两种芜菁花叶病毒株系对拟南芥的发育有不同的影响,特别是花茎的伸长,这使得病毒决定因素 P3 蛋白的表型、细胞和分子特征得以确定。两种株系的瞬时表达野生型绿色荧光蛋白标记的 P3 蛋白及其选择的突变体,揭示了它们作为内质网(ER)相关外周蛋白在沿着网流动时形成点状聚集体的行为存在重要差异。所有表达的 P3 蛋白的三维(3D)模型结构都是通过 I-TASSER 蛋白质结构预测计算构建的,这些结构用于计算蛋白质表面并绘制静电势,以表征氨基酸变化对与蛋白质相互作用以及表型和亚细胞结果相关的特征的影响。位置 279 的氨基酸是影响茎发育的主要决定因素。它还决定了表达蛋白的 ER 流动速度及其最终位置。蛋白质表面静电势的明显变化与亚细胞位置的变化相关。P3 病毒蛋白中的一个单一氨基酸决定了两种株系之间影响花茎发育的所有分析差异特征。提出了一种模型,该模型假设该蛋白在与病毒 6K2 蛋白相关的病毒复制复合物的细胞内运动中起作用。两种病毒蛋白之间的这种关联类型可能在株系之间有所不同。