Méndez-López Eduardo, Aranda Miguel A
Centro de Edafología y Biología Aplicada del Segura (CEBAS)-CSIC, Department of Stress Biology and Plant Pathology, Campus Universitario de Espinardo, Murcia, Spain.
PLoS Pathog. 2023 Oct 18;19(10):e1011732. doi: 10.1371/journal.ppat.1011732. eCollection 2023 Oct.
Cysteine oxidations play important regulatory roles during animal virus infections. Despite the importance of redox modifications during plant infections, no plant virus protein has yet been shown to be regulated by cysteine oxidation. The potexvirus pepino mosaic virus (PepMV) is pandemic in tomato crops. Previously we modeled the structure of the PepMV particle and coat protein (CP) by cryo-electron microscopy and identified critical residues of the CP RNA-binding pocket that interact with the viral RNA during particle formation and viral cell-to-cell movement. The PepMV CP has a single cysteine residue (Cys127) central to its RNA binding pocket, which is highly conserved. Here we show that the Cys127Ser replacement diminishes PepMV fitness, and that PepMV CPWT is oxidized in vivo while CPC127S is not. We also show that Cys127 gets spontaneously glutathionylated in vitro, and that S-glutathionylation blocks in vitro the formation of virion-like particles (VLPs). VLPs longer than 200 nm could be formed after in planta CPC127S overexpression, while very short and dispersed VLPs were observed after CPWT overexpression. Our results strongly suggest that the CP redox status regulates CP functions via cysteine oxidation.
半胱氨酸氧化在动物病毒感染过程中发挥着重要的调节作用。尽管氧化还原修饰在植物感染过程中很重要,但尚未有植物病毒蛋白被证明受半胱氨酸氧化调节。马铃薯X病毒属的佩佩诺花叶病毒(PepMV)在番茄作物中广泛流行。此前,我们通过冷冻电子显微镜对PepMV粒子和外壳蛋白(CP)的结构进行了建模,并确定了CP RNA结合口袋中的关键残基,这些残基在粒子形成和病毒细胞间移动过程中与病毒RNA相互作用。PepMV CP在其RNA结合口袋中有一个高度保守的半胱氨酸残基(Cys127)。在此,我们表明Cys127被丝氨酸取代(Cys127Ser)会降低PepMV的适应性,并且PepMV CPWT在体内被氧化,而CPC127S则不会。我们还表明,Cys127在体外会自发地谷胱甘肽化,并且S-谷胱甘肽化在体外会阻止病毒样颗粒(VLP)的形成。在植物中过表达CPC127S后可形成长度超过200 nm的VLP,而过表达CPWT后则观察到非常短且分散的VLP。我们的结果强烈表明,CP的氧化还原状态通过半胱氨酸氧化调节CP的功能。