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拟南芥LEC2介导的本氏烟草叶片内质网靶向外源蛋白合成增强:转录组分析的见解

AtLEC2-Mediated Enhancement of Endoplasmic Reticulum-Targeted Foreign Protein Synthesis in Nicotiana benthamiana Leaves: Insights From Transcriptomic Analysis.

作者信息

Ocampo Carolina G, Vignolles Florencia, Pombo Marina A, Colombo Maria Laura, Rosli Hernan G, Petruccelli Silvana

机构信息

Centro de Investigación y Desarrollo en Tecnología de Alimentos (CIDCA), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina.

Instituto de Fisiología Vegetal, INFIVE, CONICET, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.

出版信息

Biotechnol Bioeng. 2025 Mar;122(3):615-630. doi: 10.1002/bit.28893. Epub 2024 Nov 26.

Abstract

Many proteins used in industrial and pharmaceutical applications are typically synthesized within the secretory pathway. While yeast and mammalian cells have been engineered to enhance the production of endomembrane-targeted proteins, similar strategies in plant cells remain underexplored. This study investigates the potential of arabidopsis leafy cotyledon 2 (AtLEC2), a key regulator of seed development, to enhance the production of proteins targeted to the endoplasmic reticulum (ER) in Nicotiana benthamiana leaves. Through transient expression experiments, we demonstrate that AtLEC2 selectively increases the production of ER-targeted GUS without affecting its cytosolic variant. Moreover, leaves agroinfiltrated with AtLEC2 show a significant increase in ER-GFP accumulation compared to controls lacking AtLEC2. Transcriptomic analysis reveals that AtLEC2 promotes ribosome and chloroplast biogenesis, along with the upregulation of genes involved in photosynthesis, translation, and membrane synthesis. Notably, seed-specific poly(A) binding proteins involved in RNA stability and translation initiation, as well as 3-hydroxy-3-methylglutaryl coenzyme A reductase-linked to ER hypertrophy-are highly upregulated. This study establishes a novel connection between AtLEC2 and the enhancement of ER-targeted foreign protein synthesis, paving the way for innovative strategies in plant cellular engineering.

摘要

许多用于工业和制药应用的蛋白质通常在分泌途径中合成。虽然酵母和哺乳动物细胞已被改造以提高内膜靶向蛋白的产量,但植物细胞中的类似策略仍未得到充分探索。本研究调查了种子发育的关键调节因子拟南芥叶状子叶2(AtLEC2)在增强本氏烟草叶片中内质网(ER)靶向蛋白产量方面的潜力。通过瞬时表达实验,我们证明AtLEC2选择性地增加了ER靶向GUS的产量,而不影响其胞质变体。此外,与缺乏AtLEC2的对照相比,用AtLEC2进行农杆菌浸润的叶片显示ER-GFP积累显著增加。转录组分析表明,AtLEC2促进核糖体和叶绿体生物发生,同时上调参与光合作用、翻译和膜合成的基因。值得注意的是,参与RNA稳定性和翻译起始的种子特异性多聚腺苷酸结合蛋白,以及与内质网肥大相关的3-羟基-3-甲基戊二酰辅酶A还原酶高度上调。本研究建立了AtLEC2与增强ER靶向的外源蛋白合成之间的新联系,为植物细胞工程中的创新策略铺平了道路。

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