Genetic and Functional Interaction between the proteasome and the Primary Cilium

蛋白酶体和初级纤毛之间的遗传和功能相互作用

基本信息

批准号：
8537750
负责人：
I-Chun Tsai
金额：
$ 5.94万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8537750
关键词：
Ablation Accounting Address Affect Alleles Animal Model Architecture Bardet-Biedl Syndrome Biological Assay Biological Factors Biological Process Cell Line Cell physiology Cells Cellular Stress Cilia Ciliary Body Clinical Clinical Management Code Complex Cues Data Databases Defect Development Disease Employee Strikes Erinaceidae Evaluation Exhibits Family Genes Genetic Genetic Load Genetic Variation Goals Hereditary Disease Homeostasis Human Genetics In Vitro Knowledge Link Mediating Medical Genetics Messenger RNA Modeling Mutate Mutation Nature Organelles Paracrine Communication Patients Penetrance Pericentriolar Regions Phenotype Physiological Play Process Proteins Proteome Reporter Role Series Severities Signal Pathway Signal Transduction Signaling Molecule Stress Structure System Testing Therapeutic Ubiquitin Variant Zebrafish base ciliopathy clinically relevant cohort design improved in vivo interest kinetosome knock-down multicatalytic endopeptidase complex mutant next generation sequencing novel therapeutics outcome forecast overexpression research study sensor tool ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): The ciliopathies are a group of disorders caused by structural or functional defects in cilia and their anchoring structures, the basal bodies. Despite significant advances in identifying ciliopathy genes, the basic causative disease mechanisms are poorly understood. Recent studies have highlighted the cilium as an important sensor, leading to a host of paracrine signaling defects in ciliopathy patients and model organisms. Towards understanding the mechanism of signal transduction from the cilium, we and others have uncovered evidence that intimate a direct relationship between ciliary function and the proteasome. First, a significant fraction of the proteasome localizes to the pericentriolar region; second, several basal body proteins mutated in ciliopathies interact directly with regulatory subunits of the proteasome; finally, our recent data indicate that defects in proteasome-mediated degradation of signaling molecules such as Gli3, ?-catenin, NICD and I-?B probably underlies observed signaling defects in numerous ciliary mutants. My goal is to explore the relationship between the primary cilium and the proteasome. My hypotheses are grounded on the above facts and the following two observations: a) the proteasome is known to alter its composition under certain conditions, such as cellular stress; and b) the ciliary proteome database contains several regulatory subunits of the proteasome. Therefore, I propose to ask a) whether ciliary signaling induces compositional changes of proteasome; and b) whether mutations in ciliary components of the proteasome contribute causal and/or modifying alleles to patients with ciliopathies. To answer these questions, I will 1) investigate the composition of the proteasome upon suppression of three ciliopathy proteins (BBS4, a basal body protein; IFT88, an anterograde IFT protein; and IFT139, a retrograde IFT protein); 2) test the physiological relevance of the compositional changes by overexpressing or suppressing candidate subunits of proteasome in a series of established reporter cell lines and zebrafish models suppressed for the same set of genes; and ask whether such manipulations improve or deteriorate the previously established knock-down; 3) sequence the coding regions of all apparent ciliary components of the ubiquitin-proteasome system (UPS) in a diverse ciliopathy cohort, followed by functional evaluation of appropriate changes. These studies will contribute to the better mechanistic understanding of the relationship between ciliary function and signal transduction and will inform the genetic architecture of ciliopathies. Moreover, the potential link between proteasomal function and ciliary signaling represents a significant shift in our understanding of this organelle and opens a host of new therapeutic possibilities.

描述（由申请人提供）：纤毛病是由纤毛及其锚定结构（基底体）引起的一组疾病。尽管鉴定纤毛病基因的重大进展，基本的病因机制知之甚少。最近的研究突出了纤毛作为重要的传感器，导致纤毛病患者和模型生物的许多旁分泌信号传导缺陷。为了理解纤毛信号转导的机制，我们和其他人发现了睫状功能与蛋白酶体之间直接关系的证据。首先，蛋白酶体的很大一部分位于周围三环区域。其次，在纤毛病中突变的几种基底体蛋白与蛋白酶体的调节亚基直接相互作用。最后，我们最近的数据表明，蛋白酶体介导的信号分子的降解缺陷，例如Gli3，？ - catenin，nicd和i-？b，可能是许多纤毛突变体中观察到的信号缺陷的基础。我的目标是探索主要纤毛和蛋白酶体之间的关系。我的假设基于上述事实和以下两个观察结果：a）已知蛋白酶体在某些条件下改变其组成，例如细胞应激； b）睫状蛋白质组数据库包含蛋白酶体的几个调节亚基。因此，我建议a）睫状信号是否诱导蛋白酶体的组成变化； b）蛋白酶体的睫状成分突变是否会为纤毛病患者造成因果和/或修饰等位基因。要回答这些问题，我将1）调查蛋白酶体抑制三种纤毛病蛋白（BBS4，一种基底蛋白； IFT88，一种顺行IFT蛋白；和IFT139，逆行IFT蛋白）； 2）通过在一系列已建立的报告基因细胞系中过度表达或抑制蛋白酶体的候选亚基来测试组成变化的生理相关性，并抑制了相同基因的斑马鱼模型；并询问此类操作是改善或恶化先前确定的敲低； 3）序列在不同的纤毛病同胞组中泛素 - 蛋白酶体系统（UPS）的所有明显睫状成分的编码区域，然后对适当更改进行功能评估。这些研究将有助于更好地理解睫状功能与信号转导之间的关系，并将为纤毛病的遗传结构提供信息。此外，蛋白酶体功能与睫状信号传导之间的潜在联系代表了我们对该细胞器的理解的显着转变，并打开了许多新的治疗可能性。