RUI: Catalytic and non-catalytic targets of lysine deacetylases

该研究项目的目标是确定人类细胞中两种赖氨酸脱乙酰酶的功能，这些酶可以修饰其他蛋白质的化学结构。这些化学结构的变化改变了细胞功能；因此，赖氨酸脱乙酰酶影响许多生物过程。然而，由于与细胞内每种蛋白质的具体相互作用尚未确定，因此尚不清楚哪种赖氨酸去乙酰化酶控制哪种细胞功能以及如何控制。更好地了解赖氨酸脱乙酰酶的工作原理将增强我们对细胞生长和发育等重要生物过程是如何被调节的理解。该项目还将加强路易斯安那州泽维尔大学（Xavier University of Louisiana，一所历史上的黑人学院或大学）的研究环境，使本科生能够参与假说驱动的、高影响力的研究。参与者将接受相关的培训，以进入研究生课程和科学工作队伍。此外，研究基础设施和成果将用于支持本科生高级实验课程的培训。赖氨酸去乙酰化酶（kdac）具有酶促作用（蛋白质去乙酰化）和非酶促作用（结合成调节基因表达和蛋白质功能的复合物）。尽管人类细胞中蛋白质普遍存在乙酰化，但很少发现kdac的特定底物或间接靶点。该项目将阐明两种kdac的具体功能，以及这些功能是否涉及体内催化。鉴定两种kdac的靶蛋白；这些靶点可以直接去乙酰化和/或这些靶点可能通过其他调节功能受到影响。含有基因组编码的催化无活性kdac的细胞系将用于评估细胞内底物蛋白的乙酰化，这些将被验证以验证直接去乙酰化，评估选择性，并将体外反应速率与体内数据相关联。该项目还将确定被认为针对单个KDAC的KDAC抑制剂的细胞内特异性。总的来说，该项目将极大地扩展对KDACs在体内如何发挥作用的理解，并提供对KDACs发挥作用的不同机制和途径的见解。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The goal of this research project is to define the functions of two lysine deacetylase enzymes from human cells that modify the chemical structure of other proteins. These changes in chemical structure alter cellular functions; therefore, lysine deacetylases impact many biological processes. However, it is not known which lysine deacetylase controls which cellular function and how, because the specific interactions with each protein within cells have not yet been determined. Better knowledge of the workings of lysine deacetylases will enhance our understanding of how important biological processes such as cellular growth and development are regulated. The project will also strengthen the research environment at Xavier University of Louisiana, a Historically Black College or University, by enabling the participation of undergraduate students in hypothesis-driven, high impact research. Participants will receive training relevant for progression into graduate programs and the scientific workforce. Additionally, the research infrastructure and results will be used to support training of undergraduate students in advanced laboratory courses.Lysine deacetylases (KDACs) have both enzymatic roles (deacetylation of proteins) and non-enzymatic roles (association into complexes that regulate gene expression and protein function). Despite the widespread acetylation of proteins in human cells, few specific substrates or indirect targets of KDACs have been identified. The project will clarify the specific functions of two KDACs and whether those functions involve catalysis in vivo. The target proteins of the two KDACs will be identified; these targets could be directly deacetylated and/or these targets maybe affected via other regulatory functions. Cell lines containing genomically encoded catalytically inactive KDACs will be used to evaluate the acetylation of intracellular substrate proteins, and these will be validated to verify direct deacetylation, evaluate selectivity, and correlate in vitro reaction rates with in vivo data. The project will also determine the intracellular specificity of KDAC inhibitors believed to target a single KDAC. Overall, this project will greatly expand understanding of how KDACs function in vivo and provide insight into the different mechanisms and pathways through which KDACs exert their effects.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.