Zhai Ying, Gnanasekaran Prabu, Pappu Hanu R
Department of Plant Pathology, Washington State University, Pullman, WA 99164, USA.
Pathogens. 2021 Jan 1;10(1):27. doi: 10.3390/pathogens10010027.
Tomato spotted wilt virus (TSWV; species ) is an economically important plant virus that infects multiple horticultural crops on a global scale. TSWV encodes a non-structural protein NSs that acts as a suppressor of host RNA silencing machinery during infection. Despite extensive structural and functional analyses having been carried out on TSWV NSs, its protein-interacting targets in host plants are still largely unknown. Here, we systemically investigated NSs-interacting proteins in via affinity purification and mass spectrometry (AP-MS) analysis. Forty-three TSWV NSs-interacting candidates were identified in . Gene Ontology (GO) and protein-protein interaction (PPI) network analyses were carried out on their closest homologs in tobacco (), tomatoes () and (). The results showed that NSs preferentially interacts with plant defense-related proteins such as calmodulin (CaM), importin, carbonic anhydrase and two heat shock proteins (HSPs): HSP70 and HSP90. As two major nodes in the PPI network, CaM and importin subunit α were selected for the further verification of their interactions with NSs via yeast two-hybrid (Y2H) screening. Our work suggests that the downstream signaling, transportation and/or metabolic pathways of host-NSs-interacting proteins may play critical roles in NSs-facilitated TSWV infection.
番茄斑萎病毒（TSWV；种 ）是一种在经济上具有重要意义的植物病毒，它在全球范围内感染多种园艺作物。TSWV编码一种非结构蛋白NSs，在感染过程中作为宿主RNA沉默机制的抑制因子。尽管已经对TSWV NSs进行了广泛的结构和功能分析，但其在宿主植物中的蛋白质相互作用靶点仍 largely未知。在这里，我们通过亲和纯化和质谱（AP-MS）分析系统地研究了 中与NSs相互作用的蛋白质。在 中鉴定出了43个与TSWV NSs相互作用的候选蛋白。对它们在烟草（ ）、番茄（ ）和 中的最接近同源物进行了基因本体（GO）和蛋白质-蛋白质相互作用（PPI）网络分析。结果表明，NSs优先与植物防御相关蛋白相互作用，如钙调蛋白（CaM）、输入蛋白、碳酸酐酶和两种热休克蛋白（HSPs）：HSP70和HSP90。作为PPI网络中的两个主要节点，通过酵母双杂交（Y2H）筛选选择CaM和输入蛋白亚基α进一步验证它们与NSs的相互作用。我们的工作表明，宿主与NSs相互作用蛋白的下游信号传导、运输和/或代谢途径可能在NSs促进的TSWV感染中起关键作用。
