Molecular and Physiological Function of the Tubby Gene Family

Tubby 基因家族的分子和生理功能

基本信息

批准号：
8636456
负责人：
JUERGEN K. NAGGERT
金额：
$ 36.91万
依托单位：
JACKSON LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-09 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8636456
关键词：
Ablation Accounting Adipocytes Adipose tissue Alleles Alstrom syndrome Bardet-Biedl Syndrome Biochemical Pathway Biological Biological Process Blindness Brown Fat Cell physiology Cells Cellular Structures Centers for Disease Control and Prevention (U.S.)Central obesity Chimeric Proteins Chronic Cilia Co-Immunoprecipitations Comorbidity Complex Defect Development Disease Embryo Epidemic Eukaryota Evolution Fatty acid glycerol esters Functional disorder Gene Family Gene Proteins General Population Genes Genetic Genetic Polymorphism Goals Health Care Costs Homeostasis Human Hybrids Hypothalamic structure Image Immunoprecipitation Individual Investigation Knowledge Label Lateral Lead Leber&apos s amaurosis Maps Microscopy Microtubule-Associated Proteins Molecular Mus Mutant Strains Mice Mutate Mutation Neurons Non-Insulin-Dependent Diabetes Mellitus Obesity Organism Overweight Pathway interactions Peripheral Phenotype Photoreceptors Physiological Play Population Process Proteins Public Health Reagent Resolution Retina Retinal Retinal Diseases Retinitis Pigmentosa Rhodopsin Role Signal Transduction Site Syndrome Tamoxifen Testing Therapeutic Thermogenesis Tissues Transgenic Mice Translating Vesicle Weight Work Yeasts base body system ciliopathy disease phenotype fatty acid oxidation hearing impairment hypocretin insight mature animal member null mutation positional cloning prevent protein function protein transport public health relevance research study trafficking trend yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): We have described a small gene family, the tubby like proteins (TULP). Mutations in its founding member, tubby (TUB), cause a tripartite phenotype of obesity, retinal and cochlear degeneration in mice, which is similar to the phenotypes observed in individuals with Alstrom or Bardet-Biedl syndromes which are caused by mutations in ciliary associated proteins. The late onset, slowly progressing obesity in tubby mice is reflective of common forms of maturity onset human obesity. Polymorphisms in the TUB gene have been shown to be associated with obesity, and mutations in TULP1 cause retinitis pigmentosa and Leber's Congenital Amaurosis in human populations. Furthermore, a null mutation for Tulp3 leads to embryonic lethality in mice. A strong conservation of the tubby gene family through evolution in higher eukaryotes and the associated diseases observed when they are mutated, indicate that TULPs play an important role in the normal function of the cells in which they are expressed. Currently, the molecular function of TULPs is not known. However, accumulation of rhodopsin containing vesicles in the retina of tubby and Tulp1 mutant mice, and interaction with genes and proteins involved in intracellular trafficking, suggest a role for TULPs in this process. To begin to understand the cellular function of TUB, we will carry out experiments in genetically modified mice that will test whether TUB plays a role in cellular transport using high resolution imaging of labeled TUB protein in co-localization studies with markers of intracellular trafficking. We will also use yeast two hybrid analysis and immunoprecipitation to identify proteins that directly interact with TUB. These studies will define pathways in which TUB participates. Similar to the lack of knowledge about the cellular function of TULPs, there is also limited knowledge about how disruptions in TULPs lead to the observed disease phenotypes. To test whether the obesity in tubby mice is a result of its disruption in the CNS or in adipocytes or a combination of both, we will analyze tissue specific targeted mutations that inactivate TUB at both of these sites individually. We will test the hypothesis that tubby mice have chronic sympathetic activation to brown fat, thus limiting their obesity. We will test the hypothesis that mutations of TULP3 will lead to obesity and retinal disease in the adult animal by generating a tamoxifen inducible allele of TULP3. And finally, by positional cloning we will identify a genetic modifier of the tubby mutation that can prevent obesity in tubby mice. This suite of experiments will provide entry points into the pathways in which TULPs act and insight into their cellular function and, thereby, reveal their mechanisms of action.

描述（由申请人提供）：我们描述了一个小基因家族，例如蛋白（Tulp）。其创始成员Tubby（TUB）中的突变引起小鼠的肥胖，视网膜和人工耳蜗变性的三方表型，这与在Alstrom或Bardet-Bardet-Biedl综合征中观察到的表型相似，这些表型是由睫状相关蛋白突变引起的。较晚的发作，慢慢发展的小鼠肥胖反映了人类肥胖的常见形式。浴缸基因中的多态性已被证明与肥胖有关，TULP1的突变会导致色素性视网膜炎和列伯人群中的先天性amaurisos。此外，TULP3的无效突变导致小鼠的胚胎致死性。通过在较高的真核生物中的进化以及在突变时观察到的相关疾病的强烈保护，表明郁金香在表达其表达细胞的正常功能中起重要作用。目前，尚不清楚郁金香的分子功能。然而，在tubby和tulp1突变小鼠视网膜中含有囊泡的视紫红质的积累，以及与参与细胞内贩运的基因和蛋白质的相互作用，这表明在这一过程中塔尔普斯的作用。为了开始理解浴缸的细胞功能，我们将在转基因小鼠中进行实验，这些实验将测试浴缸在使用细胞内运输标记的共定位研究中使用高分辨率的TAUB蛋白进行高分辨率成像在细胞转运中起作用。我们还将使用酵母两种混合分析和免疫沉淀来识别直接与浴缸相互作用的蛋白质。这些研究将定义浴缸参与的途径。与缺乏有关郁金香细胞功能的知识相似，关于郁金香中断导致观察到的疾病表型的知识也有限。为了测试肥胖小鼠的肥胖是由于中枢神经系统中的破坏还是在脂肪细胞中的破坏或两者的组合，我们将分析组织特定的靶向突变，这些突变会单独地在这两个地点灭活浴缸。我们将检验以下假设：tubby小鼠对棕色脂肪具有慢性交感神经激活，从而限制了它们的肥胖症。我们将测试以下假设：Tulp3突变将通过产生塔莫莫昔芬诱导等位基因TULP3导致成人动物的肥胖和视网膜疾病。最后，通过位置克隆，我们将确定可以防止胆小小鼠肥胖症的管道突变的遗传修饰剂。这套实验套件将为塔尔普斯（Tulps）作用并深入了解其细胞功能的途径，从而揭示其作用机理。