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A Simple Cellular Model for Lipodystrophy

脂肪营养不良的简单细胞模型

基本信息

批准号：
10153463
负责人：
JOEL M GOODMAN
金额：
$ 33.37万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2023-05-31
项目状态：
已结题

项目摘要

PROJECT SUMMARY Cytosolic lipid droplets are unique among cellular organelles. These essential structures, found in nearly all eukaryotic cells, serve several functions besides fat storage, including serving as a depot for hormones and signaling molecules, providing phospholipid precursors for other organelles, innate immunity, and as a platform for assembly of pathogens. Unlike other organelles, droplets contain a single phospholipid leaflet which surrounds their hydrophobic cores. Despite the fact that they are the basis of both obesity, which affects two thirds of Americans, and lipodystrophy, an often fatal disease caused by the inability to properly store fat, there is still sketchy knowledge of their assembly. Recent progress has shown that, while droplets can form spontaneously, a few proteins and lipids are involved in regulating this process and ensuring that fully functional droplets are produced. While these components have been identified, much is unknown about their functions. This proposal seeks to test hypotheses regarding the mechanism of action of Pet10p (a yeast perilipin), seipin, and Scs3p (a Fit2 protein), and to gain molecular insight into how they function together. A driving hypothesis is that Pet10p collaborates in trans (i.e., at the bud rather than at the ER/droplet junction) with seipin, Scs3p, and diacylglycerol to promote and regulate droplet assembly. In Aim 1, focusing on Pet10p, the multiple phenotypes in PET10-deletion cells with be matched with specific domains to probe function. The basis of the stabilizing effect of Pet10p on droplets will be elucidated, and whether the same activity is responsible for its role in droplet formation will be determined. The focus of Aim 2 will be a deeper understanding the function of seipin, including mapping function to structure, control of targeting proteins to droplets, and its role in vectorial budding of droplets. Studies to elucidate structure from cryo-EM images will also be initiated. Aim 3 will focus on the collaboration of seipin, Pet10p and Scs3p on early steps of droplet budding. Whether seipin attracts Pet10p through seipin’s amino terminal will be determined, and the role of Pet10p and Scs3p on budding will be studied. The order of arrival at the nascent structure will be probed. Overall, the proposed research will elucidate important functions of the droplet assembly factors, and, for the first time, study how they interact.

项目摘要胞质脂滴是细胞器中独特的。这些基本结构，几乎在所有的真核细胞除了储存脂肪外，还具有多种功能，包括作为激素的储存库，信号分子，为其他细胞器提供磷脂前体，先天免疫，并作为平台来组装病原体与其他细胞器不同，液滴含有单个磷脂小叶，包围着它们的疏水核心。尽管事实上，他们都是肥胖的基础，这影响了两个三分之一的美国人，脂肪营养不良，一种由于无法正确储存脂肪而导致的致命疾病，对它们的组装仍知之甚少最近的进展表明，虽然液滴可以形成自发地，一些蛋白质和脂质参与调节这一过程，并确保充分产生功能液滴。虽然这些成分已经被识别出来，但对其有很多未知之处功能协调发展的该提案旨在测试关于Pet 10 p（酵母）作用机制的假设 perilipin）、seipin和Scs 3 p（一种Fit 2蛋白），并从分子水平深入了解它们如何共同发挥作用。一驱动假设是Pet 10 p反式协作（即，在芽而不是在ER/液滴连接处）与seipin、Scs 3 p和二酰基甘油一起促进和调节液滴组装。在目标1中，侧重于 PET 10-缺失细胞中的多种表型与特异性结构域相匹配以探针功能将阐明Pet 10 p对液滴稳定作用的基础，以及是否相同将确定其在液滴形成中的作用。目标2的重点将是更深入的了解seipin的功能，包括将功能映射到结构，控制靶向蛋白，液滴及其在液滴的矢量出芽中的作用。从冷冻EM图像中阐明结构的研究将也开始。目标3将重点关注seipin，Pet 10 p和Scs 3 p在液滴早期步骤上的合作发芽将确定seipin是否通过seipin的氨基末端吸引Pet 10 p，并且将确定将研究Pet 10 p和Scs 3 p对芽的影响。将探索到达新生结构的顺序。总体而言，拟议的研究将阐明液滴组装因素的重要功能，第一次，研究它们如何相互作用。