Vector-borne Virus Research Center, State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Institute of Plant Virology, College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.
Mol Plant Microbe Interact. 2020 Mar;33(3):412-422. doi: 10.1094/MPMI-08-19-0239-IA. Epub 2020 Jan 27.
Viral suppressors of RNA silencing (VSRs) are a cluster of viral proteins that have evolved to counteract eukaryotic antiviral RNA silencing pathways, thereby contributing to viral pathogenicity. In this study, we revealed that the matrix protein P4 encoded by rice stripe mosaic virus (RSMV), which is an emerging cytoplasmic rhabdovirus, is a weak RNA silencing suppressor. By conducting yeast two-hybrid, bimolecular fluorescence complementation, and subcellular colocalization assays, we proved that P4 interacts with the rice endogenous suppressor of gene silencing 3 (OsSGS3). We also determined that overexpression has no effect on transcription. However, P4 can promote the degradation of OsSGS3 via ubiquitination and autophagy. Additionally, a potato virus X-based expression system was used to confirm that P4 enhances the development of mosaic symptoms on leaves by promoting hydrogen peroxide accumulation but not cell death. To verify whether P4 is a pathogenicity factor in host plants, we generated transgenic -overexpressing rice plants that exhibited disease-related developmental defects including decreased plant height and excessive tillering. Our data suggest that RSMV-encoded P4 serves as a weak VSR that inhibits antiviral RNA silencing by targeting OsSGS3.
病毒 RNA 沉默抑制子(VSRs)是一组病毒蛋白,它们进化而来以对抗真核抗病毒 RNA 沉默途径,从而促进病毒的致病性。在这项研究中,我们揭示了由水稻条纹花叶病毒(RSMV)编码的基质蛋白 P4 是一种弱 RNA 沉默抑制剂。通过酵母双杂交、双分子荧光互补和亚细胞共定位实验,我们证明 P4 与水稻内源基因沉默抑制子 3(OsSGS3)相互作用。我们还确定 P4 的过表达对 转录没有影响。然而,P4 可以通过泛素化和自噬促进 OsSGS3 的降解。此外,我们还使用基于马铃薯病毒 X 的表达系统证实 P4 通过促进过氧化氢积累而不是细胞死亡来增强病毒在 叶片上的花叶症状的发展。为了验证 P4 是否是宿主植物中的致病性因子,我们生成了过表达 的转基因水稻植物,这些植物表现出与疾病相关的发育缺陷,包括植株高度降低和分蘖过度。我们的数据表明,RSMV 编码的 P4 作为一种弱 VSR,通过靶向 OsSGS3 抑制抗病毒 RNA 沉默。