Sirtuins and Metabolic Pathway Integration

描述(由申请人提供)：翻译后修饰是细胞适应环境变化的一种快速而有效的方式。生物学家普遍对促进我们对化学修饰发挥作用的机制的理解感兴趣，并希望更多地了解这些修饰对细胞复杂代谢网络动力学的影响。GM62203支持的这项工作的长期目标是了解sirtuin依赖的蛋白质酰化/去乙酰化系统(SDPADS)对原核细胞生理学的贡献。NIH之前对该项目的资助使我们能够进一步了解SDPADS在革兰氏阴性和革兰氏阳性细菌中的作用。我们现在处于一个很好的位置，可以对SDPADS的乙酰转移酶进行结构和功能分析，剖析SDPADS功能编码基因的表达调控，确定先前未知的基因也参与蛋白质活性的乙酰化控制，建立N-Lys蛋白乙酰化/去乙酰化在中枢代谢途径控制中的作用，并确定原核生物是否使用SDPADS通过调节转录调节因子的活性来直接调控基因的表达。我们将采取多学科的方法来回答原核细胞生理学的基本问题。这项建议的前两个目标集中在新发现的乙酰化对代谢和基因表达的影响，后两个目标集中在SDPADS酶的生化和结构特征以及编码SDPADS功能的基因的表达调控上。鉴于SDPADS的普遍存在，这一研究领域的进展将对我们理解所有细胞的生理学产生广泛的影响。公共卫生相关性快速控制蛋白质的生物活性对于细胞在不断变化的环境中生存至关重要。赖氨酸残基的翻译后修饰对基因的表达和代谢有着深远的影响。这项拟议的工作将促进我们对修饰/解调酶的作用机制的理解，这些酶组成了一个在所有形式的生命中保存的系统。编码修饰/去变性酶的基因的表达调控是拟议研究的一部分。在这个项目中，我们试图验证假定的新的蛋白质底物的系统，包括DNA结合的调节蛋白。

DESCRIPTION (provided by applicant): Posttranslational modification is a rapid and efficient way in which cells adapt to changes in their environment. Biologists at large are interested in advancing our understanding of the mechanisms through which chemical modifications exert their effects, and want to learn more about the impact that these modifications have on the dynamics of the complex metabolic network of the cell. The long-term goal of the work supported by GM62203 is to understand the contributions of the sirtuin- dependent protein acylation/deacylation system (SDPADS) to prokaryotic cell physiology. Previous NIH funding of this project allowed us to advance our understanding of the role of the SDPADS in Gram- negative and Gram-positive bacteria. We are now in an excellent position to perform structure-function analyses of the acetyltransferase of the SDPADS, to dissect the regulation of expression of the genes encoding SDPADS functions, to identify the function of previously unknown genes also involved in acetylation control of protein activity, to establish a role for N-Lys protein acetylation/deacetylation in the control of central metabolic pathways, and to determine if prokaryotes use the SDPADS to directly control gene expression by modulating the activity of transcription regulators. We will take a multidisciplinary approach to answer fundamental questions of prokaryotic cell physiology. The first two aims of this proposal focus on newly discovered effects of acetylation on metabolism and gene expression, and the last two aims focus on the biochemical and structural characterization of the SDPADS enzymes and on the regulation of expression of the genes encoding SDPADS functions. Given the ubiquity of the SDPADS, advances in this research area will have broad impact on our understanding of the physiology of all cells. PUBLIC HEALTH RELEVANCE Rapid control of biological activities of proteins is critical for the survival of cells in a changing environment. Posttranslational modifications of lysine residues have a profound effect on gene expression and metabolism. The proposed work will advance our understanding of the mechanism of function of the modifying/demodifying enzymes that comprise a system conserved in all forms of life. Regulation of expression of the genes encoding the modifying/demodifying enzymes is part of the proposed studies. In this project we seek to validate putative new protein substrates of the system, including DNA-binding regulatory proteins.