Supplement: Regulation of SOX Proteins by Methylation-dependent Proteolysis in Stem Cells and Development

补充：干细胞和发育中甲基化依赖性蛋白水解作用对 SOX 蛋白的调节

基本信息

批准号：
10810083
负责人：
HUI ZHANG
金额：
$ 1.01万
依托单位：
UNIVERSITY OF NEVADA LAS VEGAS
依托单位国家：
美国
项目类别：
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/10810083
关键词：
Alleles Animal Model Animals Anophthalmos Binding Biological Process Cells Complex DNA Methylation DNA Repair DNA Sequence Alteration DNA biosynthesis Development Doctor of Philosophy Dose Ectoderm Embryonic Development Endoderm Epigenetic Process Gene Amplification Genes Genetic Grant Growth Homeostasis Human Impairment Internships Laboratories Learning Disabilities Lysine Malignant Neoplasms Malignant neoplasm of brain Malignant neoplasm of esophagus Malignant neoplasm of lung Mesoderm Methylation Microphthalmos Motor Skills Pathway interactions Play Pluripotent Stem Cells Protein Dynamics Protein Methylation Proteins Proteolysis Regulation Research Role Schedule Seizures Syndrome TAL1 gene Tissues Transcriptional Regulation adult stem cell blastomere structure brain abnormalities embryo cell embryo tissue embryonic stem cell fetal stem cell graduate student induced pluripotent stem cell loss of function mutation malignant breast neoplasm motor learning mutant novel overexpression pluripotency protein degradation self-renewal stem cell model stem cells transcription factor ubiquitin ligase ubiquitin-protein ligase undergraduate student

项目摘要

Project Summary The CRL4 ubiquitin ligase complexes regulate many important biological processes such as DNA replication, DNA repair, transcriptional regulation, and epigenetic inheritance of DNA methylation. We recently found that CRL4 also regulates the self-renewal and pluripotency of embryonic stem cells (ESCs) through important stem cell proteins such as SOX2 (SRY-box2). SOX2 is a dose-dependent master transcriptional factor that plays a key role in regulating the self-renewal and pluripotency or multipotency of ESCs, iPSCs, and many fetal and adult stem cells. In ESCs, an increase of SOX2 promotes development into ectoderm and mesoderm lineages, while loss or reduction of SOX2 induces differentiation into endoderm and trophectoderm lineages. Even at the 4-cell embryonic stage, the heterogeneous binding of SOX2 to target genes determines the first lineage decision. In human, loss-of-function mutations on a single Sox2 allele is sufficient to cause the familial anophthalmia/microphthalmia syndrome associated with seizures, brain abnormalities, slow growth, delayed motor skills and learning disability. Conversely, gene amplification and over-expression of SOX2 are frequently associated with many poorly differentiated cancers including lung, esophagus, brain, and breast cancers. However, it remains unclear how the SOX2 protein level is regulated in various stem/progenitor cells during development and tissue homeostasis. We recently found that the protein stability of SOX2 is regulated by a novel CRL4-based proteolytic mechanism using lysine methylation as a proteolytic trigger in ESCs and other related cells. Genetic mutation of this particular CRL4 function impairs embryonic development. We propose to unravel the function and regulation of this novel and important CRL4-based proteolytic mechanism in regulating self-renewal and pluripotency of ESCs and during embryonic development. Our specific aim 1 is to define the roles of CRL4-based ubiquitin E3 ligases that target the methylated SOX2 protein for degradation. Our specific aim 2 is to determine how the levels of methylated SOX2 proteins are dynamically regulated during the self-renewal of ESCs. In our specific aim 3, we propose to examine how the function of SOX2 is regulated in animal development using specific genetic mutants defective in the methylation- dependent proteolysis pathway. Symantha Lloyd is a senior undergraduate student that is scheduled to conduct research proposed in the RO1 grant GM140185 for the summer of 2023. She is currently considering to become a PhD graduate student in Dr. Zhang’s Laboratory at UNLV.

项目概要 CRL4 泛素连接酶复合物调节许多重要的生物过程，例如 DNA DNA 甲基化的复制、DNA 修复、转录调控和表观遗传。最近发现CRL4还调节胚胎干细胞的自我更新和多能性 (ESC) 通过重要的干细胞蛋白如 SOX2 (SRY-box2) 具有剂量依赖性。主转录因子在调节自我更新和多能性或 ESC、iPSC 以及许多胎儿和成体干细胞的多能性在 ESC 中，SOX2 增加。促进外胚层和中胚层谱系的发育，同时 SOX2 的损失或减少即使在 4 细胞胚胎阶段，也能诱导分化为内胚层和滋养外胚层谱系。 SOX2 与靶基因的异质结合决定了人类的第一个谱系决定。单个 Sox2 等位基因的功能丧失突变足以导致家族性遗传无眼症/小眼症综合征与癫痫发作、大脑异常、生长缓慢、运动技能延迟和离线学习障碍、基因扩增和过度表达。 SOX2 通常与许多低分化癌症相关，包括肺癌、食道癌、然而，目前尚不清楚 SOX2 蛋白水平在癌症和乳腺癌中是如何调节的。我们最近发现，发育和组织稳态过程中的各种干/祖细胞。 SOX2 的蛋白质稳定性由使用赖氨酸的基于 CRL4 的新型蛋白水解机制调节甲基化作为 ESC 和其他相关细胞中的蛋白水解触发因素。 CRL4 功能会损害胚胎发育。这种新颖且重要的基于 CRL4 的蛋白水解机制在调节自我更新和我们的具体目标 1 是确定 ESC 的多能性和胚胎发育过程中的作用。基于 CRL4 的泛素 E3 连接酶，针对甲基化 SOX2 蛋白进行降解。目标 2 是确定甲基化 SOX2 蛋白的水平如何在在我们的具体目标 3 中，我们建议研究 SOX2 的功能。使用甲基化缺陷的特定基因突变体在动物发育中进行调节 Symantha Lloyd 是一名预定的高年级本科生。开展 2023 年夏季 RO1 拨款 GM140185 中提议的研究。她目前正在考虑成为内华达大学拉斯维加斯分校张博士实验室的博士研究生。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Lysine Methylation-Dependent Proteolysis by the Malignant Brain Tumor (MBT) Domain Proteins.

恶性脑肿瘤 (MBT) 结构域蛋白的赖氨酸甲基化依赖性蛋白水解。

DOI：
发表时间：
2024-02-13
期刊：
影响因子：
5.6
作者：
Sun, Hong;Zhang, Hui
通讯作者：
Zhang, Hui

The assembly of mammalian SWI/SNF chromatin remodeling complexes is regulated by lysine-methylation dependent proteolysis.

哺乳动物 SWI/SNF 染色质重塑复合物的组装受赖氨酸甲基化依赖性蛋白水解作用的调节。

DOI：
发表时间：
2022-11-05
期刊：
影响因子：
16.6
作者：
Guo, Pengfei;Hoang, Nam;Sanchez, Joseph;Zhang, Elaine H;Rajawasam, Keshari;Trinidad, Kristiana;Sun, Hong;Zhang, Hui
通讯作者：
Zhang, Hui

Regulation of DNA Replication Licensing and Re-Replication by Cdt1.

Cdt1 对 DNA 复制许可和再复制的调节。