Mitotic Checkpoint Regulators in Insulin Signaling

胰岛素信号传导中的有丝分裂检查点调节剂

基本信息

批准号：
9980928
负责人：
MELANIE H. COBB
金额：
$ 29.97万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-21 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9980928
关键词：
Ablation Address Affect Alleles Aneuploidy Animals Binding Biochemistry Biological Assay Blood Glucose Cell division Cell membrane Cells Cellular biology Chromosome Segregation Clathrin Clathrin Adaptors Collection Complex Development Diabetes Mellitus Diet Disease Elements Endocytosis Ensure Exhibits Fluorescence Resonance Energy Transfer Genomics Human IRS1 gene Immune checkpoint inhibitor In Vitro Insulin Insulin Receptor Insulin Resistance Knock-in Mouse Leucine Link MXD1 gene Maps Mediating Metabolic Metabolic Diseases Metabolic syndrome Metabolism Mitogen-Activated Protein Kinases Mitosis Mitotic Mitotic Checkpoint Molecular Target Mus Mutate Non-Insulin-Dependent Diabetes Mellitus Obesity Pathway interactions Patients Phenotype Physiological Physiology Proteins Publishing Recombinant Proteins Regulation Research Resistance Role Sampling Signal Transduction Syndrome Testing Time Tissues anaphase-promoting complex diabetic diabetic patient enhancer-binding protein AP-2 exosome experimental study in vivo inhibitor/antagonist insulin regulation insulin signaling liver biopsy mouse genetics mutant new therapeutic target non-diabetic novel premature prevent receptor binding reconstitution recruit single cell sequencing transcriptome sequencing

项目摘要

Project Summary Insulin signaling is critical for multiple facets of animal physiology. Its dysregulation causes insulin resistance syndromes, such as type 2 diabetes. The spindle checkpoint ensures the fidelity of chromosome segregation and guards against aneuploidy. The key spindle checkpoint proteins Mad2 and BubR1 can simultaneously bind to Cdc20, converting it from an anaphase promoting complex/cyclosome (APC/C) activator to a subunit of an APC/C-inhibitory complex called the mitotic checkpoint complex (MCC). During checkpoint inactivation, a critical inhibitor of Mad2, p31comet promotes checkpoint inactivation and timely chromosome segregation. Recently, combining approaches in mouse genetics, cell biology, biochemistry, and single-cell genomics, we have discovered a critical role of the p31comet–Mad2–BubR1 module of mitotic regulators in insulin signaling through regulating insulin receptor (IR) endocytosis. In the mouse, p31comet ablation diminishes IR at the plasma membrane prior to insulin binding and causes defective insulin signaling in multiple tissues and metabolic syndrome. Mechanistically, Mad2 directly binds to IR through a canonical Mad2-interacting motif (MIM). IR-bound Mad2 facilitates BubR1-dependent recruitment of the clathrin adaptor AP2 to IR. p31comet blocks Mad2-BubR1 association and prevents spontaneous IR endocytosis. Mad2 and BubR1 are also required for insulin-stimulated IR endocytosis. This unexpected link between mitotic regulators and insulin signaling raises several outstanding questions that we wish to address in this proposal. In Aim 1, we will further elucidate the mechanism and regulation of insulin-stimulated IR endocytosis. In particular, we will determine how the newly discovered Mad2–BubR1 mechanism cooperates with previously described mechanisms to mediate proper IR endocytosis. We will establish how these mechanisms are regulated by insulin signaling. In Aim 2, we will test the intriguing hypothesis that insulin signaling reciprocally regulates the spindle checkpoint. In preliminary results, we have created a knock-in mouse (Insr4A/4A) with mutated IR alleles (4A) deficient for Mad2 binding. IR 4A cells have a weakened spindle checkpoint. We will determine the mechanisms by which IR promotes spindle checkpoint signaling through cellular and in vitro reconstitution experiments. In Aim 3, we will define the physiological functions of the mutual regulation between IR and mitotic regulators by examining the phenotypes of the Insr4A/4A mouse. We will test whether defective IR plasma membrane localization contributes to type 2 diabetes by comparing IR localization in liver biopsies from non-diabetic and diabetic patients. Collectively, the proposed research will further clarify the mechanism and function of the unexpected link between mitotic regulators and insulin signaling, and may establish the Mad2– BubR1–AP2 module as a novel therapeutic target for treating diabetes.

项目概要胰岛素信号传导对于动物生理学的多个方面至关重要，其失调会导致胰岛素。抵抗综合症，例如 2 型糖尿病。纺锤体检查点确保了保真度。染色体分离并防止非整倍体关键纺锤体检查点蛋白 Mad2。 BubR1 可以同时与 Cdc20 结合，将其从后期促进复合物/环体 (APC/C) 激活剂，作用于称为有丝分裂的 APC/C 抑制复合物的亚基检查点复合物 (MCC) 在检查点失活过程中，Mad2 的关键抑制剂 p31comet。最近，促进检查点失活和及时染色体分离。小鼠遗传学、细胞生物学、生物化学和单细胞基因组学的方法，我们有发现有丝分裂调节因子的 p31comet–Mad2–BubR1 模块在胰岛素信号传导中的关键作用通过调节胰岛素受体 (IR) 内吞作用，在小鼠中，p31comet 消融可减少胰岛素受体 (IR) 的内吞作用。胰岛素结合之前的质膜，导致多个组织中胰岛素信号传导缺陷，从机制上讲，Mad2 通过典型的 Mad2 相互作用直接与 IR 结合。基序 (MIM) 结合 IR 的 Mad2 促进 BubR1 依赖性网格蛋白接头 AP2 向 IR 的募集。 p31comet 阻断 Mad2-BubR1 结合并防止自发 IR 内吞作用。胰岛素刺激的 IR 内吞作用也需要这种有丝分裂调节剂之间的意外联系。胰岛素信号传导提出了我们希望在本提案中解决的几个悬而未决的问题。目标1，我们将进一步阐明胰岛素刺激IR内吞作用的机制和调控。特别是，我们将确定新发现的 Mad2-BubR1 机制如何与先前描述的介导适当 IR 内吞作用的机制我们将确定这些机制如何。在目标 2 中，我们将测试一个有趣的假设：胰岛素信号相互调节纺锤体检查点在初步结果中，我们创建了一个。 IR 4A 电池具有我们将确定IR促进纺锤体的机制。在目标 3 中，我们将通过细胞和体外重建实验来定义检查点信号传导。通过检查IR和有丝分裂调节剂之间相互调节的生理功能我们将测试 Insr4A/4A 小鼠的表型是否有缺陷的 IR 质膜定位。通过比较非糖尿病和糖尿病肝活检中的 IR 定位，有助于 2 型糖尿病的发生总的来说，拟议的研究将进一步阐明糖尿病患者的机制和功能。有丝分裂调节因子和胰岛素信号之间意想不到的联系，并可能建立 Mad2- BubR1–AP2 模块作为治疗糖尿病的新型治疗靶点。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mitotic regulators and the SHP2-MAPK pathway promote IR endocytosis and feedback regulation of insulin signaling.

有丝分裂调节因子和 SHP2-MAPK 通路促进 IR 内吞作用和胰岛素信号传导的反馈调节。

DOI：
发表时间：
2019
期刊：
影响因子：
16.6
作者：
Choi, Eunhee;Kikuchi, Sotaro;Gao, Haishan;Brodzik, Karolina;Nassour, Ibrahim;Yopp, Adam;Singal, Amit G;Zhu, Hao;Yu, Hongtao
通讯作者：
Yu, Hongtao

TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling.

TP53 通过纤毛发生和音刺猬信号传导促进人类胚胎干细胞的谱系定型。

DOI：
发表时间：
2022-02-15
期刊：
影响因子：
8.8
作者：
Sivakumar, Sushama;Qi, Shutao;Cheng, Ningyan;Sathe, Adwait A;Kanchwala, Mohammed;Kumar, Ashwani;Evers, Bret M;Xing, Chao;Yu, Hongtao
通讯作者：
Yu, Hongtao

STAG2 promotes the myelination transcriptional program in oligodendrocytes.

STAG2 促进少突胶质细胞中的髓鞘化转录程序。