Kiervel Déborah, Boissinot Sylvaine, Piccini Céline, Scheidecker Danile, Villeroy Claire, Gilmer David, Brault Véronique, Ziegler-Graff Véronique
Institut de biologie moléculaire des plantes, CNRS, Université de Strasbourg, 67084 Strasbourg, France.
INRAE, Université de Strasbourg, SVQV UMR1131, 68000 Colmar, France.
Plant Physiol. 2024 Dec 23;197(1). doi: 10.1093/plphys/kiae590.
As obligate intracellular parasites, viruses depend on host proteins and pathways for their multiplication. Among these host factors, specific nuclear proteins are involved in the life cycle of some cytoplasmic replicating RNA viruses, although their role in the viral cycle remains largely unknown. The polerovirus turnip yellows virus (TuYV) encodes a major coat protein (CP) and a 74 kDa protein known as the readthrough (RT) protein. The icosahedral viral capsid is composed of the CP and a minor component RT*, arising from a C-terminal cleavage of the full-length RT. In this study, we identified Arabidopsis (Arabidopsis thaliana) ALY family proteins as interacting partners of TuYV structural proteins using yeast 2-hybrid assays and co-immunoprecipitations in planta. ALY proteins are adaptor proteins of the THO-TREX-1 complex essential to the nuclear export of mature messenger RNAs (mRNAs). Although all 4 ALY proteins colocalized with the CP and the RT protein in the nucleus upon co-expression in agro-infiltrated Nicotiana benthamiana leaves, only the CP remained nuclear and colocalized with ALY proteins in TuYV-infected cells, suggesting that the CP is an essential partner of ALY proteins. Importantly, TuYV-infected A. thaliana 4xaly knock-out mutants showed a significant increase in viral accumulation, indicating that TuYV infection is affected by an unknown ALY-mediated antiviral defense mechanism or impairs the cellular mRNA export pathway to favor viral RNA translation. This finding underpins the crucial role played by nuclear factors in the life cycle of cytoplasmic RNA viruses.
作为专性细胞内寄生虫,病毒依靠宿主蛋白和途径进行增殖。在这些宿主因子中,特定的核蛋白参与了一些在细胞质中复制的RNA病毒的生命周期,尽管它们在病毒周期中的作用仍 largely unknown。芜菁黄化病毒(TuYV)是一种粉虱传双生病毒,编码一种主要衣壳蛋白(CP)和一种74 kDa的通读(RT)蛋白。二十面体病毒衣壳由CP和一个次要成分RT组成,RT来自全长RT的C末端切割。在本研究中,我们通过酵母双杂交试验和植物体内的免疫共沉淀,将拟南芥ALY家族蛋白鉴定为TuYV结构蛋白的相互作用伴侣。ALY蛋白是THO-TREX-1复合物的衔接蛋白,对成熟信使RNA(mRNA)的核输出至关重要。尽管在农杆菌浸润的本氏烟草叶片中共表达时,所有4种ALY蛋白都与CP和RT蛋白在细胞核中共定位,但在TuYV感染的细胞中,只有CP保留在细胞核中并与ALY蛋白共定位,这表明CP是ALY蛋白的必需伴侣。重要的是,TuYV感染的拟南芥4xaly敲除突变体显示病毒积累显著增加,这表明TuYV感染受未知的ALY介导的抗病毒防御机制影响,或损害细胞mRNA输出途径以利于病毒RNA翻译。这一发现强调了核因子在细胞质RNA病毒生命周期中所起的关键作用。