Regulation of Translation by O-GlcNAc - Resubmission 03-05-2020

O-GlcNAc 翻译调节 - 重新提交 03-05-2020

• 批准号: 10308411
• 负责人: GERALD Warren HART
• 金额: $ 49.78万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10308411
• 关键词: Address; Aging; Alzheimer' s Disease; Binding; Binding Sites; Cell Nucleus; Cell physiology; Cells; Chronic Disease; Communication; Cytoplasm; Cytoplasmic Granules; Cytoplasmic Protein; DNA Modification Methylases; Data; Diabetes Mellitus; Diabetic mouse; Disease; EIF-2alpha; Eating; Enzymes; Etiology; Foundations; Gene Expression; Genetic Transcription; Genetic Translation; Glucose; Goals; Heat Stress Disorders; Histones; Hyperglycemia; In Vitro; Investigation; Kinetics; Malignant Neoplasms; Mediating; Messenger RNA; Metabolic Diseases; Methods; Mitochondrial Proteins; Modification; Molecular; Nerve Degeneration; Nuclear Proteins; Nutrient; O-GlcNAc transferase; Oryctolagus cuniculus; Peptides; Phosphorylation; Play; Polyribosomes; Post-Translational Protein Processing; Preparation; Process; Proteasome Inhibition; Protein Biosynthesis; Proteins; RNA; RNA Polymerase II; Regulation; Reticulocytes; Retina; Ribonucleoproteins; Ribosomal Proteins; Ribosomes; Role; Signal Transduction; Site; Stress; System; Time; Tissues; Toxic effect; Translational Regulation; Translations; Ubiquitination; Work; base; mRNA Decay; messenger ribonucleoprotein; multicatalytic endopeptidase complex; novel; peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase; polypeptide; premature; prevent; protein aggregation; protein expression; proteostasis; response; sensor; stress granule; sugar; transcription factor; transcriptome; transcriptome sequencing; translation factor

The cycling of N-acetylglucosamine on Ser(Thr) residues (O-GlcNAcylation; OGN) on nuclear, cytoplasmic and mitochondrial proteins serves as a nutrient sensor to regulate signaling, transcription, and cellular physiology. Abnormal OGN underlies the etiology of diabetes, cancer and Alzheimer’s disease. OGN regulates nearly every aspect of transcription, including RNA polymerase II, histones, DNA methyltransferases, and nearly all transcription factors. Recent findings by us and others indicate that O- GlcNAcylation also regulates protein translation and mRNA utilization, but much less is known. Understanding how nutrients and stress regulate protein translation via OGN is not only critical to our basic understanding of one of the cell’s most vital processes, but also is key to understanding mechanisms underlying chronic diseases of aging, such as diabetes, cancer and neurodegeneration. We hypothesize that O-GlcNAc cycling on proteins in the translational machinery regulates proteostasis by mediating communication between the proteasome and ribosomal machinery, and that nutrients regulate translation rates and mRNA selection by dynamic O-GlcNAc cycling on many ribosome-associated proteins. We propose three specific aims to advance our understanding of OGN’s roles in nutrient regulation of translation: Aim 1 will use state-of-the-art mass spectrometric methods to identify both nascent and mature ribosome associated proteins and translation factors that are modified by OGN, and we will specifically focus on those that appear to be involved in ribosome:proteasome communication. We will then determine the functions of OGN at the site level on selected OGN translation proteins. Aim 2 will elucidate how high glucose alters the OGN of the translation machinery. Using both live HEK293 cells and a rabbit reticulocyte translation system, we will determine if OGN plays a role in mRNA selection by performing RNA seq analyses of polysome preparations. Aim 3 will determine the mechanisms by which the O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) are rapidly targeted to ribosomes in response to proteasome inhibition. These studies are not only elucidating molecular mechanisms of how nutrients regulate protein synthesis, but they also are key to revealing how hyperglycemia, as occurs in diabetes, abnormally alters protein expression in many tissues. Molecular mechanisms revealed in these studies will likely lead to totally novel targets for the treatment of chronic diseases of aging, particularly diabetes.

N-乙酰氨基葡萄糖在核上 Ser(Thr) 残基上的循环（O-GlcNAcylation；OGN）， 细胞质和线粒体蛋白作为营养传感器来调节信号传导、转录和 细胞生理学异常是糖尿病、癌症和阿尔茨海默病的病因。 调节转录的几乎各个方面，包括 RNA 聚合酶 II、组蛋白、DNA 我们和其他人最近的发现表明 O-。 GlcNAcylation 还调节蛋白质翻译和 mRNA 利用，但人们知之甚少。 了解营养物质和压力如何通过 OGN 调节蛋白质翻译不仅对我们的基础知识至关重要 了解细胞最重要的过程之一，也是理解机制的关键 潜在的慢性衰老疾病，如糖尿病、癌症和神经退行性疾病。 我们研究了翻译机器中蛋白质上的 O-GlcNAc 循环调节蛋白质稳态 通过介导蛋白酶体和核糖体机器之间的通讯，并且营养物质调节 通过动态 O-GlcNAc 循环对许多核糖体相关蛋白进行翻译速率和 mRNA 选择。 我们提出了三个具体目标，以加深我们对 OGN 在营养调节中的作用的理解 翻译：目标 1 将使用最先进的质谱方法来识别新生和成熟的 经过 OGN 修饰的核糖体相关蛋白和翻译因子，我们将具体 重点关注那些似乎参与核糖体：蛋白酶体通讯的物质，然后我们将确定。 OGN 在选定的 OGN 翻译蛋白上的位点水平的功能将阐明有多高。 葡萄糖改变翻译机制的 OGN 使用活 HEK293 细胞和兔网织红细胞。 翻译系统，我们将通过进行 RNA seq 分析来确定 OGN 是否在 mRNA 选择中发挥作用 目标 3 将确定 O-GlcNAc 转移酶 (OGT) 的机制。 和 O-GlcNAcase (OGA) 响应蛋白酶体抑制而快速靶向核糖体。 这些研究不仅阐明了营养物质如何调节蛋白质合成的分子机制， 但它们也是揭示高血糖（如糖尿病中发生的）如何异常改变蛋白质的关键 这些研究揭示的分子机制可能会带来全新的结果。 治疗老年慢性疾病，特别是糖尿病的目标。