AtHVA22a是Reep/DP1/Yop1家族蛋白的植物特异性同源物,参与了芜菁花叶病毒的繁殖。

IF 4.8 1区 农林科学 Q1 PLANT SCIENCES
Mingshuo Xue, Luc Sofer, Vincent Simon, Nathalie Arvy, Mamoudou Diop, Roxane Lion, Guillaume Beucher, Amandine Bordat, Jens Tilsner, Jean-Luc Gallois, Sylvie German-Retana
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引用次数: 0

摘要

壶状病毒是最大的单链正义 RNA 病毒属,是农作物严重病害的元凶,其运动非常复杂。由于壶状病毒开发了劫持宿主分泌途径和质粒(PD)进行运输的策略,本研究的目标是鉴定与 6K2 蛋白相互作用的膜和/或 PD 蛋白,6K2 蛋白是一种壶状病毒蛋白,参与了芜菁花叶病毒(TuMV)的复制和细胞间移动。利用分裂泛素膜酵母双杂交试验,我们筛选了拟南芥 cDNA 文库,以寻找与 TuMV6K2 相互作用的蛋白。我们分离出了 AtHVA22a(脱落酸应答基因 22),它属于跨膜蛋白的多基因家族,与动物和酵母中的受体表达增强蛋白(Reep)/息肉病中删除蛋白(DP1)/Yop1 家族蛋白同源。HVA22/DP1/Yop1家族基因广泛分布于真核生物中,但HVA22蛋白在植物中的作用还不是很清楚,不过从拟南芥悬浮细胞中纯化的PD组分的蛋白质组学分析表明,AtHVA22a在PD蛋白质组中高度富集。我们利用双分子荧光互补法证实了 TuMV6K2 和 AtHVA22a 在酵母和植物体内的相互作用,并表明 TuMV6K2/AtHVA22a 的相互作用发生在 TuMV 感染期间的病毒复制区水平。最后,我们还发现,当 AtHVA22a 在植物体内过表达时,TuMV 的传播速度会加快,但通过 CRISPR-Cas9 诱变 AtHVA22a 时,TuMV 的传播速度会减慢。总之,我们的研究结果表明,AtHVA22a对TuMV的传播有促进作用,而蛋白的C端尾部在这一过程中非常重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
AtHVA22a, a plant-specific homologue of Reep/DP1/Yop1 family proteins is involved in turnip mosaic virus propagation.

The movement of potyviruses, the largest genus of single-stranded, positive-sense RNA viruses responsible for serious diseases in crops, is very complex. As potyviruses developed strategies to hijack the host secretory pathway and plasmodesmata (PD) for their transport, the goal of this study was to identify membrane and/or PD-proteins that interact with the 6K2 protein, a potyviral protein involved in replication and cell-to-cell movement of turnip mosaic virus (TuMV). Using split-ubiquitin membrane yeast two-hybrid assays, we screened an Arabidopsis cDNA library for interactors of TuMV6K2. We isolated AtHVA22a (Hordeum vulgare abscisic acid responsive gene 22), which belongs to a multigenic family of transmembrane proteins, homologous to Receptor expression-enhancing protein (Reep)/Deleted in polyposis (DP1)/Yop1 family proteins in animal and yeast. HVA22/DP1/Yop1 family genes are widely distributed in eukaryotes, but the role of HVA22 proteins in plants is still not well known, although proteomics analysis of PD fractions purified from Arabidopsis suspension cells showed that AtHVA22a is highly enriched in a PD proteome. We confirmed the interaction between TuMV6K2 and AtHVA22a in yeast, as well as in planta by using bimolecular fluorescence complementation and showed that TuMV6K2/AtHVA22a interaction occurs at the level of the viral replication compartment during TuMV infection. Finally, we showed that the propagation of TuMV is increased when AtHVA22a is overexpressed in planta but slowed down upon mutagenesis of AtHVA22a by CRISPR-Cas9. Altogether, our results indicate that AtHVA22a plays an agonistic effect on TuMV propagation and that the C-terminal tail of the protein is important in this process.

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来源期刊
Molecular plant pathology
Molecular plant pathology 生物-植物科学
CiteScore
9.40
自引率
4.10%
发文量
120
审稿时长
6-12 weeks
期刊介绍: Molecular Plant Pathology is now an open access journal. Authors pay an article processing charge to publish in the journal and all articles will be freely available to anyone. BSPP members will be granted a 20% discount on article charges. The Editorial focus and policy of the journal has not be changed and the editorial team will continue to apply the same rigorous standards of peer review and acceptance criteria.
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