Regulation of Rhythmic m6A RNA Modification by ER‐associated Degradation

ERα相关降解对节律性 m6A RNA 修饰的调节

• 批准号: 10615181
• 负责人: Deyu Fang
• 金额: $ 54.91万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10615181
• 关键词: Address; Animal Model; Apoptosis; Binding; Cellular Metabolic Process; Circadian Dysregulation; Circadian Rhythms; Cues; Cytosol; Degradation Pathway; Endoplasmic Reticulum; Enzymes; Fatty Liver; Genetic Engineering; Goals; Hepatic; High Fat Diet; Homeostasis; Human; Hyperlipidemia; Knockout Mice; Lipids; Liver; Mediating; Messenger RNA; Metabolic; Metabolic Control; Metabolic Diseases; Metabolism; Methylation; Modification; Molecular; Monitor; Mus; Pathway interactions; Periodicity; Physiological; Physiological Processes; Play; Polyubiquitination; Process; Proliferating; Protein Family; Proteins; Quality Control; RNA; Reader; Regulation; Role; Signal Transduction Pathway; Testing; Therapeutic; Time-restricted feeding; Translations; Ubiquitination; circadian; circadian pacemaker; cofactor; knock-down; lipid metabolism; mRNA Stability; mRNA Translation; metabolic phenotype; misfolded protein; non-alcoholic fatty liver disease; novel; preservation; programs; risk mitigation; ubiquitin-protein ligase

Endoplasmic Reticulum (ER)-Associated Degradation (ERAD) is a major ER quality-control program that monitors and translocates unfolded or misfolded protein substrates from the ER to cytosol for polyubiquitination and proteasomal degradation. N6-methyladenosine (m6A) methylation, the most prevalent internal modification of mammalian mRNAs, is known to regulate the stability, translation, and function of almost every major class of human RNAs. Three major families of proteins, including writers, readers, and erasers, are known to be responsible for the reversible RNA m6A methylation process. However, the signal transduction pathway underlying the regulation of RNA m6A modification remain elusive. Herein, we accumulated strong preliminary evidence for an unprecedented circadian-regulated ERAD pathway that controls mRNA m6A modification and subsequent lipid homeostasis, which we called “circadian ERAD-m6A”. Our major preliminary findings include: (i) the ER-resident E3 ubiquitin ligase HRD1 and its co-factor SEL1L, the major components of ERAD machinery, are regulated by the circadian clock in the liver; (ii) HRD1 interacts with and mediates polyubiquitination and degradation of the specific m6A writer METTL14 and the reader YTHDF3; (iii) HRD1 liver-specific KO (LKO) mice display reversed fashions with METTL14-LKO or YTHDF3-knockdown mice in hepatic m6A mRNA methylation levels, expression of lipid metabolic regulators, and metabolic phenotypes associated with hepatic steatosis and hyperlipidemia; and (iv) unlike the classic ERAD, the newly-identified ERAD-m6A regulatory axis and its function in hepatic lipid metabolism are under the control of circadian rhythm. These observations led to our central hypothesis that the liver HRD1-ERAD program, which is oscillated under the circadian clock, regulates hepatic m6A RNA modification by controlling rhythmic degradation of the specific m6A writer METTL14 and the reader YTHDF3. This unprecedented circadian ERAD-m6A RNA modification regulatory network, which may be dysregulated by circadian-disrupting cues, represents a major pathway that controls metabolic homeostasis associated with hepatic steatosis and hyperlipidemia. In this application, we will utilize molecular and cellular approaches, genetically engineered animal models, and high-throughput profiling of m6A RNA modification to critically address the function and mechanism by which circadian ERAD regulates hepatic m6A RNA modification and lipid metabolism. In two aims, we will: 1) define a novel circadian ERAD pathway that modulates rhythmic m6A RNA modification through degrading the specific m6A writer and reader; and 2) determine the functional significance of circadian ERAD-m6A RNA modification pathway in maintaining lipid homeostasis. Upon completion of this project, we will reveal the function and mechanism by which a novel circadian ERAD-m6A RNA modification pathway regulates lipid homeostasis associated with metabolic disorders. The findings will open up new paradigms for the studies on the physiological ERAD and m6A RNA modification and shed new light on developing therapeutics for metabolic disease.

内质网（ER）相关降解（ERAD）是一个主要的ER质量控制计划， 监测并易位的蛋白质底物从ER到胞质溶胶进行多泛素化 和蛋白酶体降解。 N6-甲基拉第义氨酸（M6A）甲基化，最普遍的内部修饰 众所周知，哺乳动物mRNA的众所周知可以调节几乎所有主要类别的稳定性，翻译和功能 人类RNA。众所周知，包括作家，读者和橡皮擦在内的三个主要蛋白质家族是 负责可逆的RNA M6A甲基化过程。但是，信号转导途径 RNA M6A修饰的调节基础仍然难以捉摸。在这里，我们积累了强大的初步 控制mRNA M6A修饰和 随后的脂质稳态，我们称之为“昼夜节律ERAD-M6A”。我们的主要初步发现包括： （i）ER居住的E3泛素连接酶HRD1及其联合因素SEL1L，Erad Machinery的主要组件， 由肝脏中的昼夜节律调节； （ii）HRD1与多泛素化和介导 特定M6A作者Mettl14和读取器YTHDF3的降解； （iii）HRD1肝特异性KO（LKO） 小鼠在肝M6a mRNA中显示了用mettl14-lko或ythdf3-knockdown小鼠逆转时尚 甲基化水平，脂质代谢调节剂的表达以及与肝相关的代谢表型 脂肪变性和高脂血症； （iv）与经典的Erad不同，新识别的ERAD-M6A调节轴 它在肝脂质代谢中的功能受昼夜节律的控制。这些观察导致 我们的核心假设是，在昼夜节律下振荡的肝HRD1-eRAD计划 通过控制特定M6A作者Mettl14的节奏降解来调节肝M6A RNA修饰 和读者ythdf3。这个前所未有的昼夜节律ERAD-M6A RNA修改调节网络，该网络 昼夜节律暗示可能会失去失调，代表控制代谢的主要途径 与肝脂肪变性和高脂血症相关的稳态。 在此应用中，我们将利用分子和细胞方法，基因工程动物模型， M6A RNA修饰的高通量分析，以批判性地解决该功能和机制 昼夜节律ERAD调节肝M6A RNA修饰和脂质代谢。在两个目标中，我们将：1）定义 新颖的昼夜节律途径，通过降解特定 M6A作家和读者； 2）确定昼夜节律修饰的功能意义 维持脂质稳态的途径。该项目完成后，我们将揭示功能和 新型昼夜节律ERAD-M6A RNA修饰途径调节脂质稳态的机制 与代谢疾病有关。这些发现将为有关生理学的研究开辟新的范式 ERAD和M6A RNA修饰，并为开发代谢疾病的治疗提供了新的启示。

项目成果

Inhalation Exposure to Airborne PM2.5 Induces Integrated Organelle Stress Response in the Liver.

吸入暴露于空气中的 PM2.5 会引起肝脏的综合细胞器应激反应。

• 发表时间: 2022
• 期刊: FASEB journal : official publication of the Federation of American Societies for Experimental Biology
• 作者: Zhang,Kezhong;Grossman,LawrenceI;Stemmer,PaulM;Kim,Hyunbae;Carruthers,Nicholas
• 通讯作者: Carruthers,Nicholas

CRISPR screening identifies the deubiquitylase ATXN3 as a PD-L1-positive regulator for tumor immune evasion.

• DOI: 10.1172/jci167728
• 发表时间: 2023-12-01
• 期刊: JOURNAL OF CLINICAL INVESTIGATION
• 影响因子: 15.9
• 作者: Wang, Shengnan;Iyer, Radhika;Han, Xiaohua;Wei, Juncheng;Li, Na;Cheng, Yang;Zhou, Yuanzhang;Gao, Qiong;Zhang, Lingqiang;Yan, Ming;Sun, Zhaolin;Fang, Deyu
• 通讯作者: Fang, Deyu

ATXN3 deubiquitinates YAP1 to promote tumor growth.

• 发表时间: 2023
• 期刊: American journal of cancer research
• 影响因子: 5.3
• 作者: Shengnan Wang;Kun Liu;Xiaohua Han;Yang Cheng;Emily Zhao;D. Brat;Zhaolin Sun;Deyu Fang