Essential role of the Ufm1 E3 ligase in intestinal homeostasis

Ufm1 E3 连接酶在肠道稳态中的重要作用

• 批准号: 9978762
• 负责人: HONGLIN LI
• 金额: $ 34.2万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978762
• 关键词: Ablation; Acute; Animal Model; Apoptosis; Apoptotic; Cell Death; Cell Survival; Cell physiology; Cells; Cellular biology; Chronic; Clinical; Colitis; Defensins; Development; Disease; Embryonic Development; Endoplasmic Reticulum; Enzymes; Epithelial Cells; Estrogen receptor positive; Genetic Models; Genetic study; Goals; Goblet Cells; Hematopoiesis; Hemorrhage; Homeostasis; Impairment; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestines; Knockout Mice; Lead; Malignant Neoplasms; Mediating; Messenger RNA; Mucosal Immunity; Muramidase; Organ; Outcome; Paneth Cells; Pathogenesis; Pathway interactions; Physiological; Play; Post-Translational Protein Processing; Predisposition; Protein Biosynthesis; Proteins; Role; Signal Pathway; Signal Transduction; Study models; System; Testing; Tissues; Transgenic Mice; Translations; Ubiquitin; Work; base; dysbiosis; endoplasmic reticulum stress; human disease; inhibitor/antagonist; insight; intestinal barrier; intestinal epithelium; intestinal homeostasis; ischemic injury; mRNA Transcript Degradation; mouse model; nervous system disorder; new therapeutic target; novel; protein complex; response; secretory protein; ubiquitin-protein ligase

Project Summary: Intestinal homeostasis is crucial for normal intestinal functions, and its disruption may lead to the pathogenesis of human diseases such as chronic inflammation and even cancer. Paneth cells play crucial roles in intestinal barrier function and mucosal immunity. Recent studies have demonstrated that impaired function of Paneth cells may lead to dysbiotic intestine and increased susceptibility to inflammatory bowel disease (IBD). The Ufm1 (Ubiquitin-fold modifier 1) conjugation system is a novel ubiquitin-like system that consists of Ufm1- sepcific E1 enzyme (Uba5), E2 enzyme (Ufc1) and a putative E3 enzyme (a protein complex containing Ufl1 and Ufbp1). Clinical and animal model studies implicate its potential involvement in many diseases such as neurological disorders, ischemic injury and cancer, but its overall functions and underlying mechanisms remain poorly understood. Genetic studies from others and us have recently demonstrated the essential role of this system in embryonic development and hematopoiesis, but its role in other systems remains to be defined. Ufl1 and Ufbp1, the key components of Ufm1 E3 ligase, are present in multiple tissues and organs and highly expressed in the intestine, yet their role in the gut is completely unknown. The observation of frequent gut bleeding in Ufl1 and Ufbp1 knockout mice prompted us to generate IEC (intestinal epithelial cell)-specific knockout mouse models of Ufl1 and Ufbp1. We found that IEC-specific ablation of either Ufl1 or Ufbp1 caused nearly complete loss of Paneth cells and partial loss of goblet cells, resulting in increased susceptibility to experimental colitis. Moreover, Ufbp1 ablation led to elevated ER (endoplasmic reticulum) stress, activation of the Unfolded Protein Response (UPR) and apoptosis in IECs. We also observed dramatic increase of mRNAs of secretory proteins such as lysozyme during the course of acute Ufbp1 ablation. We hypothesize that Ufbp1 and the ufmylation pathway plays a pivotal role in IRE1-mediated degradation of ER-associated mRNAs, thereby coordinating protein synthesis and ER protein load and maintaining ER homeostasis of Paneth cells. Ufbp1 deficiency impairs this degradation mechanism and leads to increased mRNAs and protein translation, resulting in elevated ER stress and Paneth cell death. To test this hypothesis, we propose three specific aims. Aim 1: To define Paneth cell-specific function of Ufbp1; Aim 2: To investigate the role of UPR in Paneth cell apoptosis induced by Ufbp1 deficiency; and Aim 3: To elucidate the mechanism of Ufbp1 ablation-induced disruption of ER homeostasis. The outcome of this project will provide critical insight into the physiological function and working mechanism of this important protein modification system in maintaining intestinal homeostasis., and have a significant impact on our understanding of Paneth cell biology, development of novel therapeutic targets for inflammatory disease, and the physiological function of the ufmylation pathway and its interplay with UPR.

项目摘要： 肠道内稳态对肠道正常功能至关重要，其破坏可能导致发病 慢性炎症甚至癌症等人类疾病。潘氏细胞在肠道免疫中发挥重要作用 屏障功能和粘膜免疫。最近的研究表明，潘氏功能受损 这些细胞可能导致肠道生态失调，并增加对炎症性肠病（IBD）的易感性。的 Ufm 1（Ubiquitin-fold modifiers 1）缀合系统是一种新型的泛素样系统，由Ufm 1- 特异性E1酶（Uba 5）、E2酶（Ufc 1）和推定的E3酶（含有Ufl 1的蛋白质复合物 UFBP1）。临床和动物模型研究暗示其可能参与许多疾病， 神经系统疾病，缺血性损伤和癌症，但其整体功能和潜在机制仍然存在 不太了解。来自其他人和我们的遗传学研究最近证明了这一点的重要作用。 它在胚胎发育和造血系统中的作用，但在其他系统中的作用仍有待确定。Ufl1 Ufbp 1和Ufbp 1是Ufm 1 E3连接酶的关键组分，存在于多种组织和器官中， 在肠道中表达，但它们在肠道中的作用完全未知。频繁肠道的观察 Ufl 1和Ufbp 1敲除小鼠的出血促使我们产生IEC（肠上皮细胞）特异性 Ufl 1和Ufbp 1基因敲除小鼠模型。我们发现IEC特异性的Ufl 1或Ufbp 1的消融导致了 潘氏细胞几乎完全丧失，杯状细胞部分丧失，导致对 实验性结肠炎此外，Ufbp 1的去除导致ER（内质网）应激升高， 未折叠蛋白反应（UPR）和细胞凋亡。我们还观察到mRNA的急剧增加， 分泌蛋白如溶菌酶在急性Ufbp 1消融过程中。我们假设Ufbp 1 并且Ufmylation途径在IRE 1介导的ER相关mRNA降解中起关键作用， 从而协调蛋白质合成和ER蛋白负载并维持潘氏细胞的ER稳态。 Ufbp 1缺陷会损害这种降解机制，并导致mRNA和蛋白质翻译增加， 导致ER应激升高和潘氏细胞死亡。为了验证这一假设，我们提出了三个具体目标。 目的1：明确Ufbp 1在潘氏细胞中的特异性功能;目的2：探讨UPR在潘氏细胞中的作用 目的3：探讨Ufbp 1基因缺失诱导细胞凋亡的机制 破坏内质网内稳态。该项目的结果将为生理学提供重要见解 这一重要的蛋白质修饰系统在维持肠道功能中的作用和工作机制 体内平衡，并对我们理解潘氏细胞生物学， 炎症性疾病的新治疗靶点和ufmylation途径的生理功能 及其与普遍定期审议的相互作用。

项目成果

DDRGK1, a crucial player of ufmylation system, is indispensable for autophagic degradation by regulating lysosomal function.

• DOI: 10.1038/s41419-021-03694-9
• 发表时间: 2021-04-20
• 期刊: Cell death & disease
• 影响因子: 9
• 作者: Cao Y;Li R;Shen M;Li C;Zou Y;Jiang Q;Liu S;Lu C;Li H;Liu H;Cai Y
• 通讯作者: Cai Y

CDK5RAP3, a Novel Nucleoplasmic Shuttle, Deeply Regulates HSF1-Mediated Heat Stress Response and Protects Mammary Epithelial Cells from Heat Injury.

CDK5RAP3 是一种新型核质梭，深度调节 HSF1 介导的热应激反应并保护乳腺上皮细胞免受热损伤

• DOI: 10.3390/ijms21218400
• 发表时间: 2020-11-09
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Shen Y;Zou Y;Li J;Chen F;Li H;Cai Y
• 通讯作者: Cai Y

DNMT3B overexpression contributes to aberrant DNA methylation and MYC-driven tumor maintenance in T-ALL and Burkitt's lymphoma.

• DOI: 10.18632/oncotarget.20176
• 发表时间: 2017-09-29
• 期刊: Oncotarget
• 作者: Poole CJ;Zheng W;Lodh A;Yevtodiyenko A;Liefwalker D;Li H;Felsher DW;van Riggelen J
• 通讯作者: van Riggelen J

ZBTB38, a novel regulator of autophagy initiation targeted by RB1CC1/FIP200 in spinal cord injury.

• DOI: 10.1016/j.gene.2018.07.073
• 发表时间: 2018-12-15
• 期刊: Gene
• 影响因子: 3.5
• 作者: Chen J;Yan L;Wang H;Zhang Z;Yu D;Xing C;Li J;Li H;Li J;Cai Y
• 通讯作者: Cai Y