pH Gradient Mitigation in the Leaf Cell Secretory Pathway Attenuates the Defense Response of Nicotiana benthamiana to Agroinfiltration.,Journal of Proteome Research

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pH Gradient Mitigation in the Leaf Cell Secretory Pathway Attenuates the Defense Response of Nicotiana benthamiana to Agroinfiltration.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-12-12 , DOI: 10.1021/acs.jproteome.9b00409
Philippe V Jutras ₁ , Frank Sainsbury ₂ , Marie-Claire Goulet ₁ , Pierre-Olivier Lavoie ₃ , Rachel Tardif ₃ , Louis-Philippe Hamel ₃ , Marc-André D'Aoust ₃ , Dominique Michaud ₁

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Partial neutralization of the Golgi lumen pH by the ectopic expression of influenza virus M2 proton channel is useful to stabilize acid-labile recombinant proteins in plant cells, but the impact of pH gradient mitigation on host cellular functions has not been investigated. Here, we assessed the unintended effects of M2 expression on the leaf proteome of Nicotiana benthamiana infiltrated with the bacterial gene vector Agrobacterium tumefaciens. An isobaric tags for relative and absolute quantification quantitative proteomics procedure was followed to compare the leaf proteomes of plants agroinfiltrated with either an "empty" vector or an M2-encoding vector. Leaves infiltrated with the empty vector had a low soluble protein content compared to noninfiltrated control leaves, associated with increased levels of stress-related proteins but decreased levels of photosynthesis-associated proteins. M2 expression partly compromised these effects of agroinfiltration to restore soluble protein content in the leaf tissue, associated with restored levels of photosynthesis-associated proteins and reduced levels of stress-related proteins in the apoplast. These data illustrate the cell-wide influence of the Golgi lumen pH homeostasis on the leaf proteome of N. benthamiana responding to microbial challenge. They also underline the relevance of assessing the eventual unintended effects of accessory proteins used to modulate specific cellular or metabolic functions in plant protein biofactories.

中文翻译：

叶细胞分泌途径中的 pH 梯度缓解减弱了本塞姆氏烟草对农杆菌渗透的防御反应。

流感病毒 M2 质子通道异位表达对高尔基体腔 pH 值的部分中和有助于稳定植物细胞中酸不稳定的重组蛋白，但 pH 梯度缓解对宿主细胞功能的影响尚未得到研究。在这里，我们评估了 M2 表达对用细菌基因载体根癌农杆菌浸润的本塞姆氏烟草叶片蛋白质组的意外影响。随后使用用于相对和绝对定量定量蛋白质组学程序的同量异位标签来比较用“空”载体或M2编码载体渗透的植物的叶蛋白质组。与未浸润的对照叶片相比，用空载体浸润的叶片具有较低的可溶性蛋白质含量，这与胁迫相关蛋白质水平增加但光合作用相关蛋白质水平降低相关。 M2 表达部分损害了农杆菌渗透恢复叶组织中可溶性蛋白含量的作用，这与光合作用相关蛋白水平的恢复和质外体中胁迫相关蛋白水平的降低有关。这些数据说明了高尔基体腔 pH 稳态对本塞姆氏烟草响应微生物挑战的叶蛋白质组的全细胞影响。他们还强调了评估用于调节植物蛋白生物工厂中特定细胞或代谢功能的辅助蛋白的最终意外影响的相关性。

更新日期：2019-12-13

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