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首页> 外文期刊>Plant Pathology >Diverse conserved domains and a positively selected site in the sugarcane streak mosaic virus P1 protein are essential for RNA silencing suppression and protein stability
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Diverse conserved domains and a positively selected site in the sugarcane streak mosaic virus P1 protein are essential for RNA silencing suppression and protein stability

机译：甘蔗条纹马赛克病毒P1蛋白中不同的保守结构域和阳性选择的位点对于RNA沉默抑制和蛋白质稳定性是必不可少的

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摘要

RNA silencing is one of the conserved antiviral mechanisms in plants, and viruses encode RNA silencing suppressors (RSS) to overcome host RNA silencing and facilitate virus infection. Sugarcane streak mosaic virus (SCSMV; species Sugarcane streak mosaic virus, genus Poacevirus, family Potyviridae) is a major causal agent of sugarcane mosaic disease in many countries in Asia, including China. In this study, we used Agrobacterium co-infiltration to show that the SCSMV P1 protein, rather than the helper component-proteinase (HC-Pro), functions as a strong RSS to suppress local RNA silencing in Nicotiana benthamiana. Mutational analysis indicated that the 15 amino acids (aa; aa 1-15) of the SCSMV P1 N-terminus were not important for RNA silencing suppression, but rather another 15 aa domain (aa 108-122) containing a conserved motif (LFR/KNKQAYIST) was essential for efficient silencing suppression by P1. In addition to the 15 aa (aa 344-358) domain in the P1 N-terminus, another 15 aa domain (aa 65-79) of P1, containing the LXKA motif and one conserved aa (D78), were associated with P1 protein stability. Furthermore, substituting the histidine (H263) residue in P1 with threonine (H263T) or alanine (H263A) also affected P1 protein stability. Notably, the H263 residue is both a positively selected site and part of the serine protease catalytic triad (HDS). Taken together, our data demonstrate that SCSMV P1, and not HC-Pro, plays a functional role in suppressing RNA silencing, and also show that some conserved motifs and a positivelyselected site in the P1 protein are associated with RSS activity and protein stability.
机译：RNA沉默是植物中保守的抗病毒机制之一，病毒编码RNA沉默抑制器（RSS）克服宿主RNA沉默并促进病毒感染。甘蔗条纹马赛克病毒（SCSMV;物种甘蔗条纹马赛克病毒，Poacevirus，家庭Potviridae）是亚洲许多国家的甘蔗马赛克病的主要因果因子，包括中国。在这项研究中，我们使用农杆菌共渗透来表明SCSMV P1蛋白，而不是辅助组分 - 蛋白酶（HC-Pro），用作抑制尼古利亚纳·宾亚植物局部沉默的强卢斯。突变分析表明，SCSMV P1 N-末端的15个氨基酸（AA; AA 1-15）对于RNA沉默抑制而言并不重要，而是另外的15AA域（AA 108-122）含有保守的主题（LFR / Knkqayist）对于P1的高效沉默抑制至关重要。除了P1 n-末端中的15AA（AA 344-358）结构域之外，含有LXKA基序和一个保守的AA（D78）的P1的另外的15AA域（AA 65-79）与P1蛋白有关稳定。此外，用苏氨酸（H263T）或丙氨酸（H263A）代替P1中的组氨酸（H263）残余物也影响P1蛋白稳定性。值得注意的是，H 2 6 3残基是正面选择的位点，部分是丝氨酸蛋白酶催化三合会（HDS）的一部分。我们的数据一起表明SCSMV P1，而不是HC-Pro，在抑制RNA沉默方面发挥作用作用，并且还表明，P1蛋白中的一些保守的基序和积极选择的位点与RSS活性和蛋白质稳定性相关。

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