Genetic analysis of FRG1 function in Drosophila

果蝇FRG1功能的遗传分析

• 批准号: 7290036
• 负责人: WILLIAM W MATTOX
• 金额: $ 16.84万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7290036
• 关键词: 4q35; Adult; Affect; Animal Model; Attention; Biochemical; Biochemical Genetics; Biochemical Pathway; Biochemistry; Biological Assay; Biological Models; Cell physiology; Collection; Complement; Complex; DNA Sequence Rearrangement; Degenerative Disorder; Development; Disease; Drosophila genus; Drosophila melanogaster; Eye; Facioscapulohumeral; Facioscapulohumeral Muscular Dystrophy; Future; Genes; Genetic; Genetic Screening; Genetic Transcription; Goals; Heterogeneous Nuclear RNA; Histology; Homologous Gene; Human; In Vitro; Knowledge; Laboratories; Lesion; Link; Modeling; Molecular; Molecular Genetics; Molecular Target; Morphology; Muscle; Muscle function; Muscular Dystrophies; Mutation; Nuclear Extract; Numbers; Organism; Pathway interactions; Phenotype; Physiological; Play; Population; Positioning Attribute; Protein Overexpression; RNA Splicing; Role; Spliceosomes; Symptoms; Syndrome; System; Testing; Therapeutic; Therapeutic Intervention; Tissues; Transgenes; Transgenic Mice; Transgenic Organisms; Work; base; compound eye; experience; fly; gene function; genetic analysis; mRNA Precursor; mouse model; novel; wasting

DESCRIPTION (provided by applicant): Facioscapulohumeral muscular dystrophy (FSHD) is a progressively debilitating syndrome that is the nation's third most common type of dystrophy, yet the development of directed therapies remains hampered by our poor understanding of its molecular basis. Recent progress using a mouse model system suggests that overexpression of the FRG1 gene is sufficient to cause a number of FSHD symptoms. Thus a promising avenue for therapeutic intervention is the reduction of FRG1 expression and/or function. Several lines of evidence suggest that FRG1 associates with the RNA splicing machinery and may alter normal splicing when overexpressed, but the specific biochemical activity of FRG1 remains unknown. In this project we will utilize the powerful genetic approaches available in the fruitfly Drosophila melanogaster to identify molecular pathways through which FRG1 functions. Using the GAL4-UAS system we will target overexpression of FRG1 specifically to the indirect flight muscle of the adult fly. The morphology and function of this muscle are easily assessed and it therefore an ideal target for genetic studies. Because FSHD has a variety of extramuscular effects we will also examine the effects of FRG1 overexpression on the Drosophila compound eye, a highly ordered tissue that is frequently exploited for analysis of gene function. Using overexpression phenotypes in these tissues we will carry out genetic screens to identify modifier mutations that reverse the harmful effects of excess FRG1 expression. Because such screens are carried out in unbiased manner they offer the potential to identify novel interacting factors that would not otherwise have been considered or investigated. As no specific biochemical activity has previously been linked with FSHD, these factors will provide new targets for strategies aimed at therapeutic inhibition of FRG1 function. In parallel studies we will exploit the extensive knowledge of RNA splicing mechanisms already developed in Drosophila to ask whether FRG1 is a required component of the spliceosome. Using an in vitro splicing system we will identify the specific steps in splicing and splicing complex assembly and function where FRG1 acts. These studies will thus provide the first genetic and biochemical analysis of FRG1 in Drosophila. By developing Drosophila as a model for FSHD this work will form a basis for future studies that will elaborate a detailed understanding of both the normal molecular function of FRG1 and the effects of its overexpression in muscle. These studies thus offer the potential to uncover novel biochemical pathways that affect muscle function and disease.

描述（由申请人提供）：面肩肱型肌营养不良症（FSHD）是一种进行性衰弱综合征，是美国第三大常见的营养不良类型，但由于我们对其分子基础的了解不足，定向治疗的发展仍然受到阻碍。使用小鼠模型系统的最新进展表明，FRG 1基因的过表达足以引起许多FSHD症状。因此，治疗干预的一个有希望的途径是减少FRG 1的表达和/或功能。一些证据表明，FRG 1与RNA剪接机制相关，并且在过表达时可能改变正常剪接，但FRG 1的特异性生化活性仍然未知。在这个项目中，我们将利用强大的遗传学方法在果蝇黑腹果蝇，以确定FRG 1功能的分子途径。使用GAL 4-UAS系统，我们将FRG 1的过表达专门针对成年苍蝇的间接飞行肌肉。这种肌肉的形态和功能很容易评估，因此它是遗传研究的理想目标。由于FSHD具有多种肌外效应，我们还将研究FRG 1过表达对果蝇复眼的影响，复眼是一种高度有序的组织，经常用于基因功能分析。使用这些组织中的过表达表型，我们将进行遗传筛选，以确定逆转过量FRG 1表达的有害影响的修饰突变。因为这样的筛选是以无偏见的方式进行的，所以它们提供了识别新的相互作用因素的可能性，否则这些因素将不会被考虑或研究。由于先前没有特定的生物化学活性与FSHD相关，这些因子将为旨在治疗性抑制FRG 1功能的策略提供新的靶点。在平行的研究中，我们将利用广泛的知识，RNA剪接机制已经在果蝇中开发，问是否FRG 1是一个必要的组成部分的剪接体。使用体外剪接系统，我们将确定剪接和剪接复合物组装和功能的具体步骤，其中FRG 1的行为。因此，这些研究将提供果蝇中FRG 1的第一个遗传和生化分析。通过开发果蝇作为FSHD的模型，这项工作将为未来的研究奠定基础，这些研究将详细了解FRG 1的正常分子功能及其在肌肉中过度表达的影响。因此，这些研究提供了发现影响肌肉功能和疾病的新生化途径的潜力。