MUTANTS WITH DEFECTS IN ENDOCYTIC FUNCTION

内吞功能缺陷的突变体

• 批准号: 2177368
• 负责人: ROCKFORD Keith DRAPER
• 金额: $ 22.1万
• 依托单位: UNIVERSITY OF TEXAS DALLAS
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-01-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2177368
• 关键词: Golgi apparatus; animal tissue; antibody formation; binding proteins; electron microscopy; endocytosis; endoplasmic reticulum; exocytosis; fluorescence microscopy; gel electrophoresis; gene complementation; immunofluorescence technique; immunoprecipitation; laboratory rabbit; lethal genes; membrane permeability; molecular cloning; morphology; protein biosynthesis; protein transport; temperature sensitive mutant; vesicle /vacuole

Endocytosis is a fundamental process in eukaryotic cells important for a variety of cellular functions ranging from the acquisition of certain nutrients, regulation of cell function, including cholesterol synthesis related to atherosclerotic plaque formation in heart disease, and response of the immune system in antibody production. The overall objective of this proposal is to better understand basic molecular events underlying steps of the endocytic process. Our approach is to isolate temperature-sensitive mutants of mammalian cells that are defective in some aspect of endocytosis and then study the mutant cells to gain information about the specific events that are defective. In specific aim one, we propose a variety of physiological and morphological studies in vivo to characterize a mutant, termed V.24.1, that has defective lysosomes at the defect. In addition, we plan an in vitro approach to investigating protein targeting in the mutant cells. Specific aim two involves studying in vitro complementation between two other mutants that are defective in endosomal acidification. In specific aim three we propose to further characterize another mutant, termed 42.2, that has an undefined lesion affecting endocytosis. Specific aim four involves using a new technique we have devised to isolate mutants expressing temperature-sensitive lesions affecting early steps of endocytic vesicle formation.

内吞作用是真核细胞中的一个基本过程，对 细胞功能的多样性，从获得某些特定的 营养素，调节细胞功能，包括胆固醇合成 与心脏病中动脉粥样硬化斑块的形成有关，以及 免疫系统在抗体产生过程中的反应。整体而言 这项建议的目的是为了更好地理解基本的分子事件 内吞过程的基本步骤。我们的方法是孤立 哺乳动物细胞中存在缺陷的温度敏感突变体 一些方面的内吞作用，然后研究突变细胞以获得 有关有缺陷的特定事件的信息。具体而言 目的第一，我们提出了各种生理和形态研究。 在活体内鉴定一种名为V.24.1的有缺陷的突变体 缺损处有溶酶体。此外，我们还计划采用体外实验方法 研究突变细胞中的蛋白质靶向。具体目标二 涉及研究另外两个突变体之间的体外互补 在内体酸化方面有缺陷。具体目标三：我们 建议进一步鉴定另一个突变体，命名为42.2，它具有 影响内吞作用的不明病变。具体目标四涉及使用 我们设计的一种新技术来分离表达基因的突变体 温度敏感性病变影响内吞小泡的早期步骤 队形。