Model Organisms

项目概述（见说明）：模式生物核心将采用两种基因可处理的系统，酵母酿酒酵母和斑马鱼D. rerio，每一种都具有独特的优势。这些实验系统将有助于解剖肾脏发育和蛋白质结构和功能的基本方面。与这些模型系统相关的实验将通过使用过去四年中从Core相关活动中产生的小分子调节剂来补充。从酵母和斑马鱼模型的独特属性以及化学调节剂的使用中产生的假设将通过其他核心继续在更高的细胞类型和生物体中进行测试。反过来，使用酵母和斑马鱼的实验提供了对更复杂系统预测的快速评估。酵母核心的目标是开发并继续利用已建立的野生型和致病蛋白的表达系统，这些蛋白通过肾脏细胞的分泌途径传递。基因组学和蛋白质组学攻击将识别影响其生物发生的因素，并建立这些因素的作用机制。为了实现这些目标，酵母核心已经创建了十几个表达系统，并为合作者提供了专业知识和工具来选择这种模式生物。Core开发的特定检测方法包括评估伴侣蛋白、泛素蛋白酶体途径、自噬和化学伴侣蛋白如何影响分泌蛋白的生物发生。斑马鱼核心将利用已建立的转基因肾脏报告系，并利用小分子筛选中已建立的自动筛选技术来识别肾脏发育和疾病的化学探针。斑马鱼核心有许多转基因系，并确定了影响肾脏发育的小分子，并确定了特定因素在肾脏发育过程中的作用。合作者将能够建立和分析新创建的斑马鱼系的结果，并进行小分子筛选。总的来说，通过与其他核心的合作，从这些互补模式生物的使用中获得的知识将得到扩展，反过来，从更复杂的系统中产生的假设可以在酵母和斑马鱼中迅速和在某些情况下得到更彻底的测试。核心将会合作

PROJECT SUMMARY (See instructions): The Model Organisms Core will employ two genetically-tractable systems, the yeast S. cerevisiae and the zebrafish D. rerio, each with distinct advantages. These experimental systems will help dissect fundamental aspects of kidney development and protein structure and function. Experiments associated with these model systems will be complemented by the use of small molecule modulators that have emerged from Core associated activities over the past four years. Hypotheses arising from the unique attributes of the yeast and zebrafish models and from the use of chemical modulators will continue to be tested in higher cell types and organisms via the other Cores. In turn, experiments using yeast and zebrafish provide rapid assessments of predictions from more complex systems. The goals of the Yeast Core are to develop and continue to utilize established expression systems for wild type and disease-causing proteins that transit the secretory pathway in kidney cells. Genomic and proteomic attacks will identify factors that impact their biogenesis, and the mechanism of action of these factors will be established. Toward these goals, the Yeast Core has created over a dozen expression systems and offers collaborators the expertise and tools to co-opt this model organism. Specific assays developed in the Core include methods to assess how chaperones, the ubiquitin proteasome pathway, autophagy, and chemical chaperones impact secretory protein biogenesis. The Zebrafish Core will utilize established transgenic kidney reporter lines and to utilize established automated screening technologies in small molecule screens to identify chemical probes for kidney development and disease. The Zebrafish Core has a number of transgenic lines and identified small molecules that impact kidney development, and has pinpointed when specific factors act during kidney development. Collaborators will be able to establish and analyze results from newly created zebrafish lines and perform small molecule screens. Overall, the knowledge gained from the use of these complementary model organisms will be expanded via collaborations with the other Cores, and in turn the hypotheses that arise from more complex systems can be rapidly and in some cases more thoroughly tested in yeast and zebrafish. The Core will co-