UBIQUITINATION LENS PROLIFERATION/DIFFERENTIATION

泛素化镜片增殖/分化

• 批准号: 6518704
• 负责人: ALLEN TAYLOR
• 金额: $ 25.2万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6518704
• 关键词: animal tissue; autoradiography; cell differentiation; cell proliferation; enzyme mechanism; gene targeting; genetically modified animals; green fluorescent proteins; growth /development; immunoprecipitation; in situ hybridization; laboratory mouse; laboratory rat; lens; northern blottings; protein isoforms; tissue /cell culture; ubiquitin; western blottings

DESCRIPTION (Adapted from applicant's abstract): During formation and growth lens cells must execute a program of proliferation, mitotic withdrawal and differentiation. In most cells, in order to progress from proliferative to differentiated phenotypes, many regulatory and structural proteins (i.e. p57) are removed via the ubiquitin (Ub) proteolytic pathway. Function of the Ub proteolytic pathway is found in all eucaryotic cells and is implicated in virtually every facet of cellular regulation, including cell cycle progression and differentiation, the stress response, DNA repair, signal transduction, gene regulation, control of protooncogenes, cellular remodeling, antigen presentation, and removal of damaged proteins. We have demonstrated the presence of a functional Ub pathway in lens cells and gathered preliminary data which is consistent with the Ub pathway regulating progress from a mitotic to a differentiated phenotype. However, the role of the Ub pathway in regulation of cell cycle and differentiation in lens has been the focus of only three publications, and no specific substrate for the Ub dependent pathway has been identified in a whole lens cell system. Based on our initial publications, preliminary data and data from other cell types, we hypothesize that the Ub proteolytic pathway, particularly ubiquitin conjugating enzymes Ubc2,3,and 4, are required to allow transition of lens cells from a proliferative to a differentiated cell type. We will test this hypothesis using lens explants and in vivo. Our long-term goals are to identify and demonstrate the function of components of the Ub proteolytic pathway, including substrates, which regulate lens cell proliferation and differentiation. The investigators have experience in all of the techniques which are required for this research. In addition, they will have 1) the guidance of Drs. Borras and Zelenka for infection of lens cells with adenovirus, and 2) the contributions of dominant negative constructs of all available Ubc and Ubc knockout mice from Drs. Pagano and Wing. Since there is only a limited literature regarding functions of the Ub pathway in mammalian systems, this work will be of interest to all scholars of mammalian development and differentiation. Dr. Taylor's group is a major contributor to the literature regarding Ub pathway functions in response to cellular stress.

描述（改编自申请人的摘要）：在形成和成长过程中 镜头细胞必须执行一个增殖，有丝分裂戒断和 分化。在大多数细胞中，为了从增殖发展到 分化的表型，许多调节和结构蛋白（即p57） 通过泛素（UB）蛋白水解途径去除。 UB的功能 蛋白水解途径均在所有胞核细胞中发现，并与 实际上，细胞调节的每个方面，包括细胞周期进程 和分化，应力响应，DNA修复，信号转导，基因 调节，原子基因的控制，细胞重塑，抗原 表现，并去除受损的蛋白质。我们已经证明了 在镜头细胞中存在功能性UB途径并收集了初步数据 这与UB途径相一致，该途径调节从有丝分裂到A的进度 分化的表型。但是，UB途径在调节中的作用 细胞周期和镜头的分化仅是三个重点 出版物，并且没有针对UB依赖途径的具体底物 在整个晶状体细胞系统中鉴定。根据我们的首次出版物， 初步数据和来自其他单元类型的数据，我们假设UB 蛋白水解途径，尤其是泛素结合酶UBC2,3和4，4， 需要允许晶状体细胞从增殖到A 分化的细胞类型。我们将使用晶状体外植体和 体内。我们的长期目标是识别和证明 UB蛋白水解途径的组成部分，包括底物，调节 透镜细胞增殖和分化。调查人员有经验 在本研究所需的所有技术中。此外， 他们将有1）Drs的指导。 Borras和Zelenka感染了镜头 带有腺病毒的细胞，以及2）主要负面构建体的贡献 在DRS的所有可用UBC和UBC淘汰小鼠中。 Pagano和Wing。自从 关于UB途径的功能，只有有限的文献 哺乳动物系统，这项工作将引起所有哺乳动物学者的兴趣 发展与分化。泰勒博士的小组是 有关UB途径响应细胞应激的文献作用。