Functional Specificity of Drosophila homeoprotein DNA binding complexes

果蝇同源蛋白 DNA 结合复合物的功能特异性

• 批准号: 7781317
• 负责人: Zenobia C Cofer
• 金额: $ 4.14万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7781317
• 关键词: Abdomen; Anterior; Behavior; Binding; Binding Proteins; Binding Sites; Cell physiology; Complex; DNA Binding; DNA-Binding Proteins; Development; Developmental Process; Diagnosis; Disease; Dissection; Drosophila genus; Ectopic Expression; Electrophoretic Mobility Shift Assay; Elements; Embryo; Enhancers; Family; Gene Expression; Gene Targeting; Genes; Genetic; Genome; Goals; Homeodomain Proteins; Human; In Vitro; Intervention; Knowledge; Lead; Learning; Life; Malignant Neoplasms; Mediating; Methods; Mutation; Nucleotides; Pattern; Process; Protein Binding; Proteins; Reporter; Repression; Role; Site; Specificity; Staging; Tertiary Protein Structure; Testing; Time; Transgenes; Transgenic Organisms; Work; cofactor; gene repression; genetic analysis; in vivo; insight; leukemia; member; mutant; novel strategies; preference; protein expression; public health relevance; research study

DESCRIPTION (provided by applicant): Developmental processes involve regulatory cascades among groups of genes that encode DMA binding proteins, which direct the full range of cellular behaviors required for normal function at all life stages. Much remains to be learned about how members of conserved families of DMA binding proteins act in concert to orchestrate patterns of gene expression. This project will investigate one of the most highly conserved of these families, the homeodomain proteins, which are known to be involved in developmental abnormalities and cancer/leukemia progression, among other important disease processes. The broad, long-term goal of these studies is to understand how DMA binding proteins interact with each other and their target sites in the genome to achieve functional specificity during development. The Engrailed homeodomain protein interacts with several other members of the same DNA binding superfamily to help it find and regulate the appropriate target genes. This proposal is to investigate underlying mechanisms using a combination of in vitro DNA binding analysis and the genetic and transgenic methods available in Drosophila, including reporter transgenes, transgenes that generate ectopic protein expression, and rescuing transgenes that mimic the endogenous pattern of expression, each used in both wildtype and mutant genetic backgrounds. The specific aims are: 1) to identify and dissect binding sites for Engrailed within an identified target enhancer in the direct target gene sloppy-paired through a combination of in vitro binding studies and in vivo transgenic analysis; and 2) to dissect the protein domains necessary for Engrailed to cooperate with its cofactors on those target sites, and test the requirements for those domains for in vivo function. PUBLIC HEALTH RELEVANCE: Fundamental knowledge of mechanisms that underlie cellular processes has proven to be essential for developing truly novel approaches to diagnosis and intervention in disease processes of all kinds, most notably in cancer development, where genetic changes accumulating over time are now understood to result in the abnormal cellular behaviors that lead to cancer. These studies will extend our understanding of how genes regulate other genes by studying a family of interacting DNA binding proteins that are highly conserved from Drosophila to humans.

描述（由申请人提供）：发展过程涉及编码DMA结合蛋白的基因组之间的调节级联，这些基因指导正常功能在所有生命阶段所需的全部细胞行为。关于DMA结合蛋白保守家族的成员如何协同编排基因表达模式，还有很多尚待了解。该项目将研究这些家族中最高度保守的同源域蛋白之一，众所周知，这些蛋白涉及发育异常和癌症/白血病进展，以及其他重要疾病过程。这些研究的广泛，长期目标是了解DMA结合蛋白如何相互相互作用及其基因组中的目标位点，以在发育过程中实现功能特异性。插入的同源域蛋白与同一DNA结合超家族的其他几个成员相互作用，以帮助其找到和调节适当的靶基因。该建议是使用体外DNA结合分析的组合以及果蝇中可用的遗传和转基因方法的组合研究的基本机制，包括报告基因转基因，转基因，产生异位蛋白质表达的转基因，并挽救模拟于表达的转型基因，每种源自野生型和突变的遗传背景。具体目的是：1）通过体外结合研究和体内转基因分析的结合，在直接靶基因sloppy中识别和剖析了构成靶向增强子中插入的结合位点； 2）剖析与其在这些目标位点上合作的辅助因子所需的蛋白质结构域，并测试这些域对体内功能的要求。公共卫生相关性：基于细胞过程的机制的基本知识已被证明对于开发真正的新颖诊断和干预各种疾病过程的方法至关重要，最值得注意的是在癌症发展中，随着时间的流逝，遗传变化随着时间的流逝而被理解为导致异常细胞行为导致癌症的异常细胞行为。这些研究将扩展我们对基因如何通过研究从果蝇到人高度保守的相互作用DNA结合蛋白的家族来调节其他基因的理解。