Model Organisms

项目摘要(见说明)：模式生物核心将使用两个遗传易处理的系统，酵母S.cerevisiae和斑马鱼D.rerio，每个系统都有不同的优势。这些实验系统将有助于剖析肾脏发育以及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核心相关活动中出现的小分子调节器来补充。由酵母和斑马鱼模型的独特属性以及化学调节剂的使用而产生的假设将继续通过其他核心在高等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的预测的快速评估。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进行传递。基因组和蛋白质组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增选这种模式生物。核心中开发的具体分析包括评估伴侣、泛素蛋白酶体途径、自噬和化学伴侣如何影响分泌蛋白生物发生的方法。斑马鱼核心将利用已建立的转基因肾脏报告系，并利用已建立的小分子筛查中的自动筛选技术来识别肾脏发育和疾病的化学探针。斑马鱼核心拥有许多转基因品系，并确定了影响肾脏发育的小分子，并准确地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品系的结果，并进行小分子筛选。总体而言，从使用这些互补的模式生物获得的知识将通过与其他核心的合作而得到扩展，反过来，从更复杂的系统中产生的假设可以迅速地在酵母和斑马鱼中得到更彻底的测试。核心将共同-

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-