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: 6171678
• 负责人: ROBERT W BRACKENBURY
• 金额: $ 23.25万
• 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/6171678
• 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 这个修改后的应用是上皮细胞运动被抑制 E-钙粘蛋白介导的细胞间接触产生的信号。 首席研究员实验室发表的研究结果表明 E-钙粘蛋白的连环蛋白结合域是粘附所必需的，但不是 抑制运动，而 JM 结构域（包含 30-50 个氨基酸 就在质膜旁边）抑制运动但不支持 许多细胞类型中的粘附。 来自新合作者 Michael Kinch 的数据， 表明E-钙粘蛋白的参与刺激酪氨酸 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，然后确定是哪一种 需要 E-cad 尾部的一部分来生成该酪氨酸 磷酸化反应并鉴定酪氨酸磷酸化蛋白。 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方面的作用。 最终的目标是 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 在培养物中表达 E-钙粘蛋白和 N-钙粘蛋白的显性失活形式 细胞，然后使用 K14 启动子靶向选定的显性失活 构建到转基因小鼠的表皮上。