Regulation of caveolae internalization

描述(由申请人提供)： 这项拟议的研究将有助于阐明通过小凹调节内吞作用的分子机制和调控元件。具体地说，将使用体外小凹萌发试验来确定肌动蛋白和小窝蛋白-1在稳定细胞表面小凹方面的作用。此外，发芽试验和柱层析将被用来鉴定小窝发芽所需的细胞质因子。将通过RNAi敲除和体外实验来研究PKC(α)和血清剥夺反应蛋白在调节小窝萌发中的作用。此外，还将研究配体结合对小窝内化受体的影响。所有使用体外试验确定的因子都将被测试它们在整个细胞中的相关性。许多致病因素，包括细菌(例如衣原体)、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降解的途径。因此，更深入地了解小窝萌发的调控机制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.