Elucidating the functions of RPD3/HDA1-like lysine deacetylases in Arabidopsis thaliana

赖氨酸ε-氨基的可逆乙酰化最近被发现是一种普遍存在的翻译后修饰，控制着转录以外的许多重要细胞功能。它是一种发生在原核生物和真核生物蛋白质上的保守修饰，表明它对生命的普遍重要性。许多代谢酶上赖氨酸乙酰化位点的鉴定是揭示其在代谢中的生理作用的第一步。赖氨酸乙酰化是由赖氨酸乙酰转移酶和赖氨酸去乙酰化酶的拮抗作用催化的。赖氨酸去乙酰化酶（Lysine deacetylases, kdac）在调节植物的逆境防御和发育中具有核心功能，但迄今为止对其作用的研究主要是在组蛋白的背景下进行的，其分子活性的多样性仍未得到充分的探索。本项目旨在全面研究拟南芥RPD3/ hda1样kdac的分子功能，揭示与植物逆境生理发育相关的新功能和靶蛋白。该项目将使用一个复杂的生化工具箱来解剖植物中新的KDAC功能。工具箱将包括基于活性的探针，用于从组织中提取活性KDAC，并通过定量质谱分析，大规模的KDAC突变体乙酰酶谱分析，以及测试新的基于肽的酰基赖氨酸底物以确定其酶特性。综上所述，这些方法将揭示新的KDAC功能对植物逆境反应和发育的整合很重要。

Reversible acetylation of the ε-amino group of lysine has recently emerged as an ubiquitous post-translational modification controlling many important cellular functions beyond transcription. It is a conserved modification occurring on proteins of both, prokaryotes and eukaryotes, indicating its universal importance for life. The identification of lysine acetylation sites on many metabolic enzymes was a first step to uncover its physiological role in metabolism. Lysine acetylation is catalysed by the antagonistic action of lysine acetyltransferases and lysine deacetylases. Lysine deacetylases (KDACs) have central functions in regulating stress defenses and development in plants, but their action was mainly investigated in the context of histones thus far, and their versatility in molecular activities is still largely unexplored. This project aims at a comprehensive investigation on the molecular functions of Arabidopsis RPD3/HDA1-like KDACs to uncover new functions and target proteins connected to plant stress physiology and development. This project will employ a sophisticated biochemical toolbox for dissecting novel KDAC functions in plants. The toolbox will include activity-based probes for the pull-down of active KDACs from tissues and analysed by quantitative mass spectrometry, large-scale acetylome profiling of KDAC mutants, and testing new peptide-based acyl-lysine substrates to determine their enzymatic properties. Taken together these approaches shall uncover novel KDAC functions important for the integration of stress responses and development in plants.