Cucurbit Chlorotic Yellows Virus p22 Protein Interacts with Cucumber SKP1LB1 and Its F-Box-Like Motif Is Crucial for Silencing Suppressor Activity
Cucurbit Chlorotic Yellows Virus p22 Protein Interacts with Cucumber SKP1LB1 and Its F-Box-Like Motif Is Crucial for Silencing Suppressor Activity
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葫芦褪绿黄化病毒 p22 蛋白与黄瓜 SKP1LB1 相互作用，其 F-Box 样基序对于沉默抑制子活性至关重要
DOI：
10.3390/v11090818
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发表时间：
2019-09-01
期刊：
影响因子：
4.7
通讯作者：
Shi, Yan
中科院分区：
文献类型：
--
作者：
Chen, Siyu;Sun, Xinyan;Shi, Yan
植物利用RNA沉默来防御病毒。作为回应，病毒编码各种RNA沉默抑制子来抵消抗病毒沉默。在这里，我们发现p22是葫芦绿黄病毒的沉默抑制因子，并发现p22与黄瓜s期激酶相关蛋白1 （SKP1）的同源物CsSKP1LB1相互作用。通过序列分析确定了p22的f -box样基序，并通过酵母双杂交试验发现了相互作用所必需的。确定了F-box-like motif参与p22沉默抑制子活性。对表达p22及其f -box样基元缺失突变体的烟叶进行蛋白质组学分析，发现228个差异表达蛋白和5个富集的京都基因与基因组百科（KEGG）通路：ABC转运蛋白、倍半萜类和三萜类生物合成、泛素介导的蛋白质水解、核黄素代谢、半胱氨酸和蛋氨酸代谢。总之，我们的研究结果证明了p22和CsSKP1LB1之间的相互作用，并表明F-box-like motif的缺失抑制了p22沉默抑制子的活性。通过蛋白质组学分析确定了p22通过F-box-like motif调控的可能途径。
Plants use RNA silencing as a defense against viruses. In response, viruses encode various RNA silencing suppressors to counteract the antiviral silencing. Here, we identified p22 as a silencing suppressor of cucurbit chlorotic yellows crinivirus and showed that p22 interacts with CsSKP1LB1, a Cucumis sativus ortholog of S-phase kinase-associated protein 1 (SKP1). The F-box-like motif of p22 was identified through sequence analysis and found to be necessary for the interaction using a yeast two-hybrid assay. The involvement of the F-box-like motif in p22 silencing suppressor activity was determined. Proteomics analysis of Nicotiana benthamiana leaves expressing p22, and its F-box-like motif deletion mutant showed 228 differentially expressed proteins and five enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways: ABC transporters, sesquiterpenoid and triterpenoid biosynthesis, ubiquitin-mediated proteolysis, riboflavin metabolism, and cysteine and methionine metabolism. Collectively, our results demonstrate the interaction between p22 and CsSKP1LB1 and show that the deletion of F-box-like motif inhibits p22 silencing suppressor activity. The possible pathways regulated by the p22 through the F-box-like motif were identified using proteomics analysis.