Carolina G Ocampo, Florencia Vignolles, Marina A Pombo, Maria Laura Colombo, Hernan G Rosli, Silvana Petruccelli
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引用次数: 0
摘要
许多用于工业和制药的蛋白质通常是在分泌途径中合成的。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。本研究调查了拟南芥多叶子叶 2(ATLEC2)(种子发育的关键调控因子)在提高拟南芥叶片内质网(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 能选择性地提高 ER 靶向 GUS 的产量,而不影响其细胞质变体。此外,与缺乏 AtLEC2 的对照组相比,灌注 AtLEC2 的叶片显示 ER-GFP 积累显著增加。转录组分析表明,AtLEC2 促进了核糖体和叶绿体的生物发生,并上调了参与光合作用、翻译和膜合成的基因。值得注意的是,与 RNA 稳定性和翻译起始有关的种子特异性多聚(A)结合蛋白以及与 ER 肥大有关的 3-羟基-3-甲基戊二酰辅酶 A 还原酶都高度上调。这项研究在 AtLEC2 与增强 ER 靶向外来蛋白合成之间建立了新的联系,为植物细胞工程的创新策略铺平了道路。
AtLEC2-Mediated Enhancement of Endoplasmic Reticulum-Targeted Foreign Protein Synthesis in Nicotiana benthamiana Leaves: Insights From Transcriptomic Analysis.
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.
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