Lysine Malonylation and SIRT5 in Epigenetic Regulation

表观遗传调控中的赖氨酸丙二酰化和 SIRT5

• 批准号: 9198466
• 负责人: Eric M. Verdin
• 金额: $ 3.3万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9198466
• 关键词: Acetyl-CoA Carboxylase; Acetylation; Active Sites; Aging; Antibodies; Bacteria; Binding; Biological; Biological Assay; Biological Process; CD4 Positive T Lymphocytes; Candidate Disease Gene; Carnitine O-Palmitoyltransferase; Cell Nucleus; Cells; Cellular Stress Response; ChIP-seq; Charge; Chromatin Structure; Cultured Cells; Cytosol; Data Set; Deacetylase; Desire for food; Diabetes Mellitus; Disease; Drops; Epigenetic Process; Family member; Fasting; Fatty Acids; Follow-Up Studies; Gene Abnormality; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Genomic Segment; Genomics; Glucose; Hepatocyte; Histone Code; Histone H1; Histone H1(s); Histone H2B; Histones; Human; Hypothalamic structure; Knockout Mice; Knowledge; Light; Link; Liver; Lysine; Malonates; Malonyl Coenzyme A; Maps; Mediating; Metabolic; Metabolic Diseases; Metabolic syndrome; Metabolism; Mitochondria; Modeling; Modification; Molecular Conformation; Mus; Nicotinamide adenine dinucleotide; Nuclear; Nutritional; Nutritional status; Obesity; Pathologic; Pattern; Physiological; Proteins; Proteomics; Regulation; Reporting; Research Proposals; Role; SIRT1 gene; Sirtuins; Site; Surveys; Testing; Tissues; Transcription Initiation Site; Transcriptional Regulation; base; epigenetic regulation; fatty acid biosynthesis; fatty acid metabolism; fatty acid oxidation; fatty acid transport; feeding; histone modification; insulin secretion; knockout gene; malonyl-CoA decarboxylase; normal aging; novel; overexpression; oxidation; response; transcriptome; transcriptome sequencing

PROJECT ABSTRACT Epigenetic factors have emerged as crucial players in metabolic disorders and in aging, both of which are typically associated with a wide range of gene expression changes. The overall objective of this proposal is to investigate the roles of a novel histone modification, lysine malonylation, and its regulation by the NAD+- dependent protein deacylase SIRT5, as a novel histone modifier in epigenetic regulation of gene expression in response to metabolic changes. The model is based on our recent identification of histone H2B lysine 5 (H2BK5) as a site of malonylation regulated by SIRT5. We propose that the dynamic malonylation of histone H2B by cellular malonyl-CoA and demalonylation by SIRT5 regulates chromatin structure and gene transcription. Two specific aims are developed to test this model globally in mouse liver and then mechanistically in cultured cells. First, genomic regions bound with malonylated histones (H2BK5) and SIRT5 will be compared using ChIP-seq using tissues from wild type and Sirt5-/- mice. These studies will aim to identify the sites of SIRT5-mediated histone demalonylation. The functional consequences of histone malonylation will be evaluated by comparing gene transcriptional changes using RNA-seq between wild type and Sirt5-/- mice. This data set will be compared to the sites where SIRT5 binds to the genome and demalonylates histones to identify the direct genomic sites of SIRT5 action. Second, we will study in mechanistic details how histone malonylation is dynamically regulated by SIRT5 and by fluctuations in cellular malonyl-CoA that occur during feeding and fasting in mice. In primary cultured mouse hepatocytes, we will also test the effect of feeding malonate (which is converted into malonyl-CoA intracellularly) and the effect of manipulating the cellular synthesis or degradation of malonyl-CoA via acetyl-CoA carboxylase (synthesis) and malonyl-coA decarboxylase (degradation). This research proposal takes the first step towards understanding the function of this newly discovered histone modification, malonylation, and its eraser, SIRT5, in epigenetic regulation. It will contribute significantly to our longstanding effort of unveiling the “histone code”, its intersection with intermediary metabolism, and to advance our knowledge of gene expression regulation by epigenetic factors. Results from the proposed project will therefore permit mechanistic follow-up studies of the significance of histone malonylation and SIRT5 in epigenetic regulation and how dysregulation may contribute to both pathological conditions, such as metabolic syndrome and diabetes, and to normal aging.

项目摘要 表观遗传因素已成为代谢性疾病和衰老的关键参与者 通常与广泛的基因表达变化有关。该提议的总体目的是 研究一种新型组蛋白修饰，赖氨酸误导性及其对NAD+ - 调节的作用 依赖性蛋白脱酰基酶SIRT5，是一种新型组蛋白修饰剂，在基因表达的表观遗传调节中 对代谢变化的反应。该模型基于我们最近对组蛋白H2B赖氨酸5的鉴定 （H2BK5）作为由SIRT5调节的误导地点。我们建议组蛋白的动态误犬 H2B通过细胞丙二酰辅-COA和SIRT5的销售量调节染色质结构和基因 转录。开发了两个具体目标，以在小鼠肝脏中全球测试该模型，然后 在培养细胞中机械上。首先，基因组区域与丙二酰化组蛋白（H2BK5）和SIRT5结合 使用芯片seq使用野生型和SIRT5 - / - 小鼠的组织进行比较。这些研究将旨在 识别SIRT5介导的Hisstone命令的地点。组蛋白的功能后果 将通过使用RNA-Seq在野生型之间比较基因转录变化来评估误习 和SIRT5 - / - 小鼠。将将此数据集与SIRT5与基因组结合的站点进行比较 将组蛋白划分为识别SIRT5作用的直接基因组位点。其次，我们将学习 机械详细说明了Hisstone Malonylation如何通过SIRT5和细胞波动动态调节 在小鼠的喂养和禁食期间发生的丙二酰coA。在原发性培养的鼠标肝细胞中，我们也将 测试丙二这酸盐的效果（细胞内转化为丙二酰辅酶A）和 通过乙酰辅酶A羧化酶（合成）和 malonyl-COA脱羧酶（降解）。 该研究建议迈出了了解这个新发现的组蛋白功能的第一步 在表观遗传调节中的修饰，误导及其橡皮sirt5。它将为我们的 长期以来揭示“组蛋白代码”，与中间代谢的交集以及 通过表观遗传因素提高我们对基因表达调节的了解。拟议项目的结果 因此，将允许对组蛋白误导和SIRT5的重要性的机械随访研究 表观遗传调节以及失调可能导致两种病理状况，例如代谢 综合征和糖尿病，以及正常衰老。