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一种紫黄质深氧化酶与一种病毒 RNA 沉默抑制剂相互作用,从而抑制病毒扩增。

A Violaxanthin Deepoxidase Interacts with a Viral Suppressor of RNA Silencing to Inhibit Virus Amplification.

机构信息

State Key Laboratory of Agrobiotechnology and Key Laboratory of Pest Monitoring and Green Management-MOA, China Agricultural University, Beijing 100193, China.

Department of Pomology/Laboratory of Stress Physiology and Molecular Biology for Tree Fruits-Key Laboratory of Beijing Municipality, China Agricultural University, Beijing 100193, China.

出版信息

Plant Physiol. 2017 Dec;175(4):1774-1794. doi: 10.1104/pp.17.00638. Epub 2017 Oct 11.

Abstract

RNA silencing plays a critical role against viral infection. To counteract this antiviral silencing, viruses have evolved various RNA silencing suppressors. Meanwhile, plants have evolved counter-counter defense strategies against RNA silencing suppression (RSS). In this study, the violaxanthin deepoxidase protein of maize (), ZmVDE, was shown to interact specifically with the helper component-proteinase (HC-Pro; a viral RNA silencing suppressor) of (SCMV) via its mature protein region by yeast two-hybrid assay, which was confirmed by coimmunoprecipitation in cells. It was demonstrated that amino acids 101 to 460 in HC-Pro and the amino acid glutamine-292 in ZmVDE mature protein were essential for this interaction. The mRNA levels of were down-regulated 75% to 65% at an early stage of SCMV infection. Interestingly, ZmVDE, which normally localized in the chloroplasts and cytoplasm, could relocalize to HC-Pro-containing aggregate bodies in the presence of HC-Pro alone or SCMV infection. In addition, ZmVDE could attenuate the RSS activity of HC-Pro in a specific protein interaction-dependent manner. Subsequently, transient silencing of the gene facilitated SCMV RNA and coat protein accumulation. Taken together, our results suggest that ZmVDE interacts with SCMV HC-Pro and attenuates its RSS activity, contributing to decreased SCMV accumulation. This study demonstrates that a host factor can be involved in secondary defense responses against viral infection in monocot plants.

摘要

RNA 沉默在抗病毒感染中起着至关重要的作用。为了对抗这种抗病毒沉默,病毒进化出了各种 RNA 沉默抑制子。与此同时,植物也进化出了针对 RNA 沉默抑制(RSS)的反防御策略。在这项研究中,通过酵母双杂交试验显示,玉米的紫黄质脱环氧化酶蛋白(ZmVDE)可通过其成熟蛋白区域与(SCMV)的辅助成分蛋白酶(HC-Pro;一种病毒 RNA 沉默抑制子)特异性相互作用,这在 细胞中通过共免疫沉淀得到了证实。研究表明,HC-Pro 中的氨基酸 101 到 460 和 ZmVDE 成熟蛋白中的谷氨酰胺 292 氨基酸对于这种相互作用是必需的。在 SCMV 感染的早期阶段,的 mRNA 水平下调了 75%至 65%。有趣的是,ZmVDE 通常定位于叶绿体和细胞质中,但在单独存在 HC-Pro 或 SCMV 感染的情况下,它可以重新定位到含有 HC-Pro 的聚集体中。此外,ZmVDE 可以以特定的蛋白质相互作用依赖的方式减弱 HC-Pro 的 RSS 活性。随后,瞬时沉默 基因促进了 SCMV RNA 和外壳蛋白的积累。综上所述,我们的研究结果表明,ZmVDE 与 SCMV HC-Pro 相互作用,并减弱其 RSS 活性,导致 SCMV 积累减少。本研究表明,宿主因子可以参与单子叶植物中针对病毒感染的次级防御反应。

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