CONTROL OF EPITHELIAL CELL MOTILITY BY E CADHERIN

E 钙粘蛋白对上皮细胞运动的控制

• 批准号: 2636112
• 负责人: ROBERT W BRACKENBURY
• 金额: $ 18.27万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-10 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2636112
• 关键词: MDCK cell; cadherins; cell motility; epithelium; genetically modified animals; keratinocyte; laboratory mouse; mammary epithelium; phosphorylation; protein structure function; wound healing; yeast two hybrid system

DESCRIPTION: (Adapted from the applicant's abstract) - The hypothesis of this revised application is that epithelial cell motility is suppressed by signals generated as a result of E-cadherin mediated cell-cell contact. Published findings from the principal investigator's lab have shown that the catenin binding domain of E-cadherin is required for adhesion, but not for suppression of motility, whereas the JM domain (comprising 30-50 amino acids just next the to plasma membrane) suppresses motility but does not support adhesion in many cell types. Data from a new collaborator, Michael Kinch, indicate that engagement of E-cadherin stimulates the tyrosine phosphorylation of several proteins in breast epithelial cells and MDCK cells. A new specific aim (Aim 1) based on these data is to determine whether this is the case for keratinocytes as well and then determine which parts of the E-cad tail are required to generate this tyrosine phosphorylation response and identify tyrosine-phosphorylated proteins. Aim 2 proposes to identify components that interact with the JM domain of E-cadherin and test their role in suppression of motility. The final aim is to determine the role of the JM domain in regulating the motility of keratinocytes in vitro and in vivo. This will be accomplished first by expressing dominant-negative forms of E-cadherin and N-cadherin in cultured cells, and then using the K14 promoter to target selected dominant-negative constructs to the epidermis of transgenic mice.

描述：（改编自申请人的摘要）-假设 该修改的应用是上皮细胞运动性被抑制， 由于E-钙粘蛋白介导的细胞-细胞接触而产生的信号。 主要研究者实验室发表的研究结果表明， E-cadherin的catenin结合结构域是粘附所必需的，但不是粘附所必需的。 抑制运动性，而JM结构域（包含30-50个氨基酸 紧邻质膜）抑制运动性，但不支持 在许多细胞类型中的粘附。 来自新合作者迈克尔·金奇的数据， 表明E-钙粘蛋白参与刺激酪氨酸 乳腺上皮细胞和MDCK中几种蛋白质的磷酸化 细胞 基于这些数据的一个新的具体目标（目标1）是确定 角质形成细胞是否也是如此，然后确定 E-cad尾的一部分是产生这种酪氨酸所必需的 磷酸化反应和鉴定酪氨酸磷酸化蛋白质。 目的 2建议识别与JM域交互的组件， E-钙粘蛋白，并测试其在抑制运动中的作用。 最终目标是 以确定JM结构域在调节运动性中的作用， 角质形成细胞在体外和体内。 这将首先通过以下方式实现： 在培养细胞中表达显性负性形式的E-钙粘蛋白和N-钙粘蛋白 细胞，然后使用K14启动子靶向选定的显性阴性 将构建体植入转基因小鼠的表皮。