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-乙酰葡萄糖在Ser（THR）固定（O-Glcnacylation; OGN）上的N-乙酰葡萄糖的循环， 细胞质和线粒体蛋白是一种营养传感器，可调节信号传导，转录和 细胞生理。异常OGN是糖尿病，癌症和阿尔茨海默氏病的病因的基础。 OGN 调节转录的几乎每个方面，包括RNA聚合酶II，组蛋白，DNA 甲基转移酶，几乎所有转录因子。我们和其他人的最新发现表明O- Glcnacylation还调节蛋白质的翻译和mRNA利用率，但知之甚少。 了解营养和压力如何调节通过OGN的蛋白质翻译不仅对我们的基本至关重要 了解细胞最重要的过程之一，但这也是理解机制的关键 型糖尿病，癌症和神经退行性等衰老的基本慢性疾病。 我们假设在转化机械中蛋白质上的O-GlCNAC循环调节蛋白质 通过介导蛋白酶体和核糖体机械之间的通信，并调节营养 通过在许多与核糖体相关蛋白上的动态O-GlCNAC循环进行的翻译速率和mRNA选择。 我们提出了三个具体的目标，以促进我们对OGN在营养调节中的作用的理解 翻译：AIM 1将使用最先进的质谱方法来识别新生和成熟 核糖体相关的蛋白质和翻译因子被OGN修饰，我们将特别具体 专注于似乎参与核糖体的：蛋白酶体通信。然后我们将确定 OGN在选定的OGN翻译蛋白上的位点级别的功能。 AIM 2将阐明有多高 葡萄糖改变了翻译机械的OGN。使用活的HEK293细胞和兔网状细胞 翻译系统，我们将通过执行RNA SEQ分析来确定OGN是否在mRNA选择中起作用 多层制剂。 AIM 3将确定O-GLCNAC转移酶（OGT）的机制 O-Glcnacase（OGA）响应蛋白酶体抑制而迅速针对核糖体。 这些研究不仅阐明了营养如何调节蛋白质合成的分子机制， 但是它们也是揭示高血糖的关键，如糖尿病中发生的那样，绝对改变了蛋白质 在许多组织中的表达。这些研究中揭示的分子机制可能会导致完全新颖 治疗慢性疾病的靶标，尤其是糖尿病。