首页> 外文期刊>Frontiers in Plant Science >Nicotiana benthamiana Elicitor-Inducible Leucine-Rich Repeat Receptor-Like Protein Assists Bamboo Mosaic Virus Cell-to-Cell Movement
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Nicotiana benthamiana Elicitor-Inducible Leucine-Rich Repeat Receptor-Like Protein Assists Bamboo Mosaic Virus Cell-to-Cell Movement

机译: Nicotiana benthamiana 诱导子诱导的富含亮氨酸的重复受体样蛋白协助竹叶病毒的细胞间运动

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For successful infection, a virus requires various host factors at different stages such as translation, targeting, replication, and spreading. One of the host genes upregulated after Nicotiana benthamiana infection with Bamboo mosaic virus (BaMV), a single-stranded positive-sense RNA potexvirus, assists in viral movement. To understand how this host protein is involved in BaMV movement, we cloned its full-length cDNA by rapid amplification of cDNA ends. The gene has 3199 nt and encodes a 969-amino acid polypeptide. The sequence of the encoded polypeptide is orthologous to that of N. tabacum elicitor-inducible leucine-rich repeat (LRR) receptor-like protein ( NtEILP ), a plant viral resistance gene, and is designated NbEILP . To reveal how NbEILP is involved in BaMV movement, we fused green fluorescent protein (GFP) to its C-terminus. Unfortunately, the gene’s expression in N. benthamiana was beyond our detection limit possibly because of its large size (~135 kDa). However, NbEILP at such low expression could still enhance BaMV accumulation in inoculated leaves. A short version of NbEILP was constructed to remove the LRR domain, NbEILP/ΔLRR-GFP; the expression of this deletion mutant could still enhance BaMV accumulation to 1.7-fold that of the control. Hence, the LRR domain in NbEILP is not an essential element in BaMV movement. We constructed a few deletion mutants — NbEILP/ΔLRRΔTMD (without the transmembrane domain), NbEILP/ΔLRRΔCD (without the cytoplasmic domain), and NbEILP/ΔLRRΔSP (without the signal peptide) — to examine whether these domains are involved in BaMV movement. For BaMV movement, NbEILP requires the signal peptide to target the endoplasmic reticulum and the transmembrane domain to retain on the membrane.
机译:为了成功感染病毒,病毒需要处于不同阶段的各种宿主因素,例如翻译,靶向,复制和传播。烟草本氏菌感染竹花叶病毒(Basm花叶病毒)(BaMV)是一种单链正向RNA马铃薯过表达病毒后,其上调的宿主基因之一有助于病毒运动。为了了解该宿主蛋白如何参与BaMV运动,我们通过快速扩增cDNA末端克隆了其全长cDNA。该基因具有3199 nt,并编码969个氨基酸的多肽。编码的多肽的序列与烟草烟草诱导子诱导的富含亮氨酸的重复序列(LRR)受体样蛋白(NtEILP)(植物病毒抗性基因)同源,并命名为NbEILP。为了揭示NbEILP如何参与BaMV运动,我们将绿色荧光蛋白(GFP)融合到其C末端。不幸的是,该基因在本氏烟草中的表达超出了我们的检测极限,可能是因为其大小较大(约135 kDa)。但是,如此低表达的NbEILP仍可增强接种叶片中BaMV的积累。构建了短版本的NbEILP以去除LRR结构域NbEILP /ΔLRR-GFP;该缺失突变体的表达仍可以使BaMV积累增加至对照的1.7倍。因此,NbEILP中的LRR域不是BaMV运动中必不可少的元素。我们构建了一些缺失突变体-NbEILP /ΔLRRΔTMD(无跨膜结构域),NbEILP /ΔLRRΔCD(无胞质结构域)和NbEILP /ΔLRRΔSP(无信号肽),以检查这些结构域是否参与BaMV运动。对于BaMV运动,NbEILP要求信号肽靶向内质网,而跨膜结构域则保留在膜上。

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