Engineered AIM-based selective autophagy to degrade proteins and organelles in planta.
Engineered AIM-based selective autophagy to degrade proteins and organelles in planta.
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基于 AIM 的工程选择性自噬可降解植物中的蛋白质和细胞器。
DOI：
10.1111/nph.18557
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Faqiang Li
中科院分区：
文献类型：
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作者：
Na Luo;Dandan Shang;Zhiwei Tang;Jinyan Mai;Xiao Huang;Li;Linchuan Liu;Caiji Gao;Yangwen Qian;Qingjun Xie;Faqiang Li
●蛋白质靶向技术是生物学研究的基本方法。最近在哺乳动物细胞中通过利用自然降解机制开发的蛋白质敲除工具允许精确测定蛋白质功能和发现降解剂型药物。然而，目前在植物中没有直接靶向内源性蛋白质进行降解的方法。●在这里，我们描述了一种新的靶向蛋白清除方法，通过工程化自噬受体与结合剂提供靶标特异性和ATG 8结合基序（AIM）将靶标连接到新生的自噬体，从而利用自噬机制进行降解。●我们通过使用基于烟草的瞬时表达系统降解各种荧光标记的蛋白质，包括胞质mCherry，核定位的bZIP转录因子TGA 5和质膜锚定的油菜素类固醇受体BRI1，以及荧光包被的过氧化物酶体，证明了其特异性和广泛的潜力。AIM自噬受体在拟南芥中的稳定表达进一步证实了这种方法在内源蛋白选择性自噬中的可行性。●由于其广泛的底物范围和特异性，我们基于AIM的工程化选择性自噬概念可以为操纵植物中的内源蛋白提供方便和强大的研究工具，并可能为植物研究开辟一条降解蛋白质以外的细胞质组分的途径。
● Protein-targeting technologies represent essential approaches in biological research. Protein knockdown tools developed recently in mammalian cells by exploiting natural degradation mechanisms allow for precise determination of protein function and discovery of degrader-type drugs. However, no method to directly target endogenous proteins for degradation is currently available in plants. ● Here, we describe a novel method for targeted protein clearance by engineering an autophagy receptor with a binder to provide target specificity and an ATG8-binding motif (AIM) to link the targets to nascent autophagosomes, thus harnessing the autophagy machinery for degradation. ● We demonstrate its specificity and broad potentials by degrading various fluorescence-tagged proteins, including cytosolic mCherry, the nucleus-localized bZIP transcription factor TGA5, and the plasma membrane-anchored brassinosteroid receptor BRI1, as well as fluorescence-coated peroxisomes, using a tobacco-based transient expression system. Stable expression of AIM-based autophagy receptors in Arabidopsis further confirms the feasibility of this approach in selective autophagy of endogenous proteins. ● With its wide substrate scope and its specificity, our concept of engineered AIM-based selective autophagy could provide a convenient and robust research tool for manipulating endogenous proteins in plants and may open an avenue toward degradation of cytoplasmic components other than proteins for plant research.