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.

项目摘要 CRL4泛素连接酶复合物调节许多重要的生物学过程，例如DNA DNA甲基化的复制，DNA修复，转录调控和表观遗传。我们最近发现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，例如SOX2（SRY-Box2）。 Sox2是剂量依赖性的主转录因素在调节自我更新和多能性方面起关键作用 ESC，IPSC以及许多胎儿和成人干细胞的多稳定性。在ESC中，Sox2的增加促进对外胚层和中胚层谱系的发展，而SOX2的损失或减少诱导分化为内胚层和滋养剂谱系。即使在4细胞胚胎阶段， SOX2对靶基因的异质结合决定了第一个谱系决策。在人类中单个Sox2等位基因上的功能丧失突变足以引起家庭与癫痫发作，脑异常，缓慢生长，性质/摩托律综合征相关的性质/粒细胞病综合征延迟运动技能和学习障碍。相反，基因扩增和过表达 Sox2经常与许多分化较差的癌症有关，包括肺，食道，大脑和乳腺癌。但是，目前尚不清楚如何调节Sox2蛋白水平发育和组织稳态期间各种茎/祖细胞。我们最近发现 Sox2的蛋白质稳定性由新型的基于CRL4的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。该特定的基因突变 CRL4功能会损害胚胎发育。我们建议阐明这种新颖而重要的基于CRL4的蛋白水解机制在调节自我更新和 ESC和胚胎发育期间的多能力。我们的特定目的1是定义基于CRL4的泛素E3连接酶，靶向甲基化的SOX2蛋白降解。我们的具体 AIM 2是确定如何在期间动态调节甲基化SOX2蛋白的水平 ESC的自我更新。在我们的特定目标3中，我们建议研究Sox2的功能在动物发育中使用特定的遗传突变体在甲基化中有缺陷依赖性蛋白水解途径。 Symantha Lloyd是一名预定的高级本科生进行2023年夏季RO1 Grant GM140185的研究。她目前是考虑成为UNLV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）

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

DOI：
10.1038/s41467-022-34348-9
发表时间：
2022-11-05
期刊：
NATURE COMMUNICATIONS
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.