Fat1 and Conventional Kinesin in Lateral Polarity

Fat1 和传统驱动蛋白的横向极性

基本信息

批准号：
8287172
负责人：
DUNCAN BRUCE JOHNSTONE
金额：
$ 5.88万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2012-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8287172
关键词：
Accounting Actins Adult Basic Science Biological Models Biology Birth Cadherins Carrier Proteins Cell Culture Techniques Cell Polarity Cells Cellular biology Cessation of life Conflict (Psychology)Data Development Disease Progression End stage renal failure Epithelial Cells Family Flowers Focal Segmental Glomerulosclerosis Foot Process Goals Health Health Expenditures Hemodialysis Integral Membrane Protein Intercellular Junctions Kidney Kidney Diseases Kinesin Knock-out Knockout Mice Lateral Left Light Literature Microscopy Modeling Molecular Biology Morbidity - disease rate Morphology Mus NPHS2 protein National Institute of Diabetes and Digestive and Kidney Diseases Nephrotic Syndrome Organ Patients Phenotype Principal Investigator Process Protein Biochemistry Proteins Proteinuria Public Health Regulation Renal glomerular disease Reporting Research Right-On Role Techniques Testing Tissues Training Transgenic Mice United States Wild Type Mouse genetic regulatory protein glomerular basement membrane improved injured intracellular protein transport mortality nephrin nephrogenesis outreach podocyte prevent protein function protein protein interaction protein transport research study scaffold slit diaphragm vasodilator-stimulated phosphoprotein

项目摘要

DESCRIPTION (provided by applicant): Fat1 is large transmembrane protein of the cadherin super-family that localizes to the podocyte slit diaphragm. Deletion of fat1 in experimental mice results in abnormal development of the slit diaphragm, massive proteinuria, and death shortly after birth with nephrotic syndrome, a phenotype similar to mice deleted for nphs1 (Nephrin). This proposal includes background and preliminary data showing that Fat1 is required for the fundamental process of lateral polarity in epithelial cells, includes preliminary data that Fat1 interacts with conventional kinesin, and hypothesizes that the interaction between Fat1 and kinesin is critical for lateral polarity. I propose a model to explain why Fat1 -dependent lateral polarity is fundamentally important for podocyte development and function, which I will test using cultured podocytes and transgenic mice. This proposal also includes background on the regulation of actin dynamics by Fat1, which may occur through interaction with the actin regulatory protein VASP. During the course of this research, I will use techniques of protein biochemistry and cell biology, and I will receive training in the use of advanced microscopy and mouse transgenics. Fat1 research has applications to general cell biology, including cell polarity, intracellular protein transport, and protein-protein interactions. Fat1 research also directly applies to the path biology of glomerular disease due to its localization at the slit diaphragm and the severe phenotype of experimentalFat1 knockout mice. PUBLIC HEALTH RELEVANCE: With governmental and professional outreach, the public is becoming increasingly aware that glomerular disease is a major cause of morbidity, mortality, and public health expenditure, accounting for over 55% of patients starting hemodialysis in the United States. Our treatments for patients with glomerular disease to slow or prevent progression of disease are not sufficiently effective, and do not precisely target the causes of glomerular disease. This proposal seeks to extend our understanding of glomerular disease through the study of slit diaphragm protein Fat1. My goal is to understand the biology ofFat1 in a simplified cell culture model system, and then to extend these studies to the role of Fat1 in the glomerulus. This proposal is relevant to the goals of the NIDDK to promote research on basic kidney biology, including applications within Cell and Molecular Biology of the Kidney (protein trafficking and cell polarity) and within the Pathobiology of Kidney Disease (normal biology of glomerular cells, and proteinuria).

描述（由申请人提供）： Fat1 是钙粘蛋白超家族的大型跨膜蛋白，定位于足细胞裂隙隔膜。实验小鼠中删除 fat1 会导致裂隙发育异常、大量蛋白尿，并在出生后不久因肾病综合征而死亡，这种表型与删除 nphs1（Nephrin）的小鼠类似。该提案包括背景和初步数据，表明 Fat1 是上皮细胞横向极性基本过程所必需的，包括 Fat1 与传统驱动蛋白相互作用的初步数据，并假设 Fat1 和驱动蛋白之间的相互作用对于横向极性至关重要。我提出了一个模型来解释为什么 Fat1 依赖性横向极性对于足细胞的发育和功能至关重要，我将使用培养的足细胞和转基因小鼠来测试该模型。该提案还包括 Fat1 调节肌动蛋白动力学的背景，这可能是通过与肌动蛋白调节蛋白 VASP 相互作用而发生的。在这项研究过程中，我将使用蛋白质生物化学和细胞生物学技术，并将接受使用先进显微镜和小鼠转基因技术的培训。 Fat1 研究可应用于一般细胞生物学，包括细胞极性、细胞内蛋白质运输和蛋白质-蛋白质相互作用。由于 Fat1 定位于裂隙隔膜以及实验性 Fat1 敲除小鼠的严重表型，Fat1 研究也直接适用于肾小球疾病的路径生物学。公共卫生相关性：随着政府和专业人士的宣传，公众越来越意识到肾小球疾病是发病、死亡和公共卫生支出的主要原因，在美国，肾小球疾病占开始血液透析患者的 55% 以上。我们对肾小球疾病患者减缓或预防疾病进展的治疗方法不够有效，并且不能精确针对肾小球疾病的病因。该提案旨在通过研究裂隙隔膜蛋白 Fat1 来扩展我们对肾小球疾病的理解。我的目标是在简化的细胞培养模型系统中了解 Fat1 的生物学特性，然后将这些研究扩展到 Fat1 在肾小球中的作用。该提案与 NIDDK 的目标相关，即促进基础肾脏生物学研究，包括肾脏细胞和分子生物学（蛋白质运输和细胞极性）和肾脏疾病病理学（肾小球细胞的正常生物学和细胞极性）中的应用。蛋白尿）。