Regulation of beta-catenin proteolysis in dorsal-ventral patterning

背腹模式中β-连环蛋白水解的调节

• 批准号: 8113749
• 负责人: ZHENGLUN ZHU
• 金额: $ 8.91万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-15 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8113749
• 关键词: 26S proteasome; Biochemical Genetics; Cell physiology; Cells; Complex; Congenital Abnormality; Data; Disease; Dorsal; Embryo; Embryonic Development; Equilibrium; Foundations; Future; Generations; Germ Layers; Goals; In Vitro; Investigation; Malignant Neoplasms; Mediating; Mesenchymal; Messenger RNA; Methods; Modeling; Molecular; Organ; Pattern; Pattern Formation; Phosphorylation; Phosphotransferases; Play; Principal Investigator; Proteins; Proteolysis; Regulation; Research; Research Personnel; Role; Signal Transduction; Stem cells; Testing; Tissues; Ubiquitin-mediated Proteolysis Pathway; Work; Xenopus; base; beta catenin; in vivo; insight; loss of function; mutant; neoplastic; novel; prevent; research study; tissue regeneration

DESCRIPTION (Provided by Applicant): The long-term goal of the proposed studies is to define the role of ubiquitin-mediated proteolysis (UMP) in cell fate determination during dorsoventral patterning. UMP by the 26S proteosome has been shown to play critical roles in regulating cellular activities. While broadly implicated in cell fate determination during early embryogenesis, currently little is known about the exact role and regulation of UMP in dorsoventral patterning, which forms the foundation for the generation of germ layers and ontogenesis of tissues and organs. Previous studies have shown that both ¿-catenin from the dorsal signaling center and Xom from the ventral signaling center are controlled by UMP. It is well appreciated that during early embryogenesis, ventral (Xom) signaling antagonizes dorsal (¿-catenin) signaling; nevertheless, the intrinsic function of UMP in dorsoventral signaling is not clear. The goal of this application is to define the function of UMP in balancing dorsal and ventral signals. The research derives from the principal investigator's preliminary studies, which allowed him to formulate the hypothesis that Xom antagonizes dorsal signaling through GSK-mediated proteolysis of ¿-catenin. Using combined biochemical and genetic approaches and a Xenopus model, the investigators propose to further test their hypothesis through the following two specific aims. Aim 1 will determine the mechanisms of Xom-induced proteolysis of ¿-catenin in vitro by defining the involvement of GSK3 kinase and Ser33/37 phosphorylation. In addition they will identify the critical domain of Xom required for inducing proteolysis of ¿-catenin. Aim 2 will define the effects of Xom on UMP of ¿-catenin in vivo by determining the effects of Xom and GSK3 in the temporal and spatial expression pattern of the ¿-catenin protein and mRNA during early embryogenesis. The results of the proposed studies will introduce a new paradigm underlying the formation of dorsal-ventral asymmetry, and are expected to pave the way for understanding stem cell function and future management of congenital malformations and neoplastic diseases. PROJECT NARRATIVE: Molecular mechanisms controlling the formation of dorsal-ventral axis formation represent a fundamental challenge of early embryogenesis. The proposed studies will focus on the role of ubiquitin-mediated proteolysis in dorsoventral patterning formation. The results of these studies will be broadly implicated in preventing congenital malformation, regulating stem cell function, as well as managing neoplastic diseases.

描述（由申请人提供）：拟议研究的长期目标是确定泛素介导的蛋白水解（UMP）在背腹侧模式过程中细胞命运决定中的作用。由26S蛋白体介导的UMP已被证明在调节细胞活动中起关键作用。虽然UMP广泛参与早期胚胎发生过程中细胞命运的决定，但目前对其在背腹侧模式中的确切作用和调控知之甚少，而背腹侧模式是胚层产生和组织器官个体发生的基础。先前的研究表明，来自背侧信号中心的¿-catenin和来自腹侧信号中心的Xom都受UMP的控制。众所周知，在胚胎发生早期，腹侧（Xom）信号可以拮抗背侧（¿-catenin）信号；然而，UMP在背腹侧信号传导中的内在功能尚不清楚。本应用的目的是确定UMP在平衡背侧和腹侧信号中的功能。这项研究源于首席研究员的初步研究，这使他能够提出Xom通过gsk介导的catenin蛋白水解拮抗背侧信号传导的假设。通过结合生化和遗传方法以及爪蟾模型，研究人员提出通过以下两个具体目标进一步验证他们的假设。目的1将通过确定GSK3激酶和Ser33/37磷酸化的参与来确定xom诱导的体外catenin蛋白水解的机制。此外，他们将确定诱导-catenin蛋白水解所需的Xom的关键结构域。Aim 2将通过测定Xom和GSK3对胚胎发生早期¿-catenin蛋白和mRNA的时空表达模式的影响，确定Xom对体内¿-catenin UMP的影响。这些研究的结果将引入背腹不对称形成的新范式，并有望为理解干细胞功能和未来先天性畸形和肿瘤疾病的治疗铺平道路。