Regulation of caveolae internalization

描述（由申请人提供）： 这项研究将有助于阐明胞饮作用的分子机制和调控元件。具体而言，将使用体外小窝出芽测定来确定肌动蛋白和小窝蛋白-1在稳定细胞表面小窝中的作用。此外，出芽试验和柱层析将用于鉴定小窝出芽所需的胞质因子。PKC（alpha）和血清剥夺反应蛋白在调节小窝出芽中的作用将使用RNAi敲低和体外测定来研究。此外，将研究配体结合对通过小窝的受体内化的影响。将检测使用体外试验鉴定的所有因子在全细胞中的相关性。许多致病因子包括细菌（例如，沙眼衣原体）病毒（例如，埃博拉和马尔堡丝状病毒）和细菌毒素（例如，霍乱毒素）通过小窝内化，从而绕过导致溶酶体中降解的途径。因此，更深入地了解小窝出芽的调控可能会导致临床应用。

DESCRIPTION (provided by applicant): The proposed research will help elucidate the molecular mechanisms and regulatory elements governing endocytosis via caveolae. Specifically, an in vitro caveolae budding assay will be used to determine the roles of actin and caveolin-1 in stabilizing caveolae at the cell surface. Furthermore, the budding assay and column chromatography will be used to identify the cytosolic factors required for caveolae budding. The roles of PKC (alpha) and the serum deprivation response protein in regulating caveolae budding will be investigated using RNAi knockdown and the in vitro assay. Furthermore, the influence of ligand binding on receptor internalization by caveolae will be investigated. All factors identified using the in vitro assay will be tested for their relevance in whole cells. Many disease-causing agents including bacteria (e.g. Chlamidia trechomatis) viruses (e.g. the Ebola and Marburg filoviruses), and bacterial toxins (e.g. cholera toxin) are internalized through caveolae, thus bypassing pathways that lead to degradation in lysosomes. Therefore, a deeper understanding of the regulation of caveolae budding could lead to clinical applications.