Indentification of Hox target genes controlling tissue growth

鉴定控制组织生长的 Hox 靶基因

• 批准号: 7753632
• 负责人: Matthew Slattery
• 金额: $ 3.2万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2010-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7753632
• 关键词: Accounting; Binding; Binding Sites; Biology; Cell Proliferation; Cell surface; Cells; Complement; DNA; DNA Binding; DNA Sequence; DNA-Binding Proteins; Development; Disease; Drosophila genome; Drosophila genus; Drosophila melanogaster Proteins; Epidermal Growth Factor; Erinaceidae; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Glypican; Goals; Grant; Growth; Homeodomain Proteins; Human; Location; Malignant Neoplasms; Methods; Microarray Analysis; Modeling; Monitor; Morphogenesis; Morphology; Organ; Organ Size; Pathway interactions; Pattern; Property; Proteins; Regulation; Regulator Genes; Relative (related person); Research; Role; Signal Pathway; Signaling Molecule; Site; Specificity; Tissues; Validation; Wing; Work; base; chromatin immunoprecipitation; cofactor; extracellular; fascinate; follow-up; genome-wide; imaginal disc; in vivo; insight; member; monomer; morphogens; novel; progenitor; public health relevance; receptor; research study; transcription factor

DESCRIPTION (provided by applicant): Disruption of tissue growth and patterning is a common step in the development of cancer, and proper tissue morphology depends upon combined input from intercellular signaling pathways and region-specific transcription factors. In Drosophila melanogaster the protein Ultrabithorax (Ubx), a region-specific transcription factor and member of Hox family of homeodomain proteins, restricts growth of hindwing (haltere) tissue in part by modulating the expression of several components of the mitogenic Decapentaplegic (Dpp) pathway. The goal of this project is to identify target genes and transcriptional regulatory elements in the Drosophila genome that are directly regulated by Ubx in the haltere; three experimental approaches will be used to accomplish this goal. First, a novel gene expression profiling .method will be used to analyze regional gene expression differences in serially homologous Ubx-expressing (haltere) and non-expressing (wing) tissues. The second step will use chromatin immunoprecipitation and DamID in combination with Drosophila genome tiling arrays to determine the DNA regions that are directly bound by Ubx in the haltere. Together, these experiments will allow the identification of direct Ubx target genes in the haltere on a genome-wide scale. The third step in this analysis will be to use Cognate Site Identity (CSI) microarray technology to characterize the DNA-binding specificity of Ubx, both as a monomer and in conjunction with the Hox cofactors Extradenticle (Exd) and Homothorax (Hth). This will permit exploration of the DNA sequences to which Ubx can and cannot bind, and, in combination with the first two steps, should allow a better understanding of a Hox regulatory network. Follow-up analysis on a subset of Ubx target genes, particularly those likely to be involved in growth regulation, will include the validation of their expression patterns in both the wing and haltere and functional analysis using standard Drosophila genetic approaches. Public Health Relevance: The Hox proteins and many of the signaling molecules controlling growth, proliferation and morphogenesis are conserved between Drosophila and humans; the goal of this research is to provide insight into the mechanisms by which a Hox transcription factor regulates tissue growth. Ultimately, a detailed description of how Hox proteins regulate tissue growth during normal development will be crucial to understanding their role when growth is misregulated in diseases states such as cancer.

描述（由申请人提供）：组织生长和模式的破坏是癌症发展的常见步骤，正确的组织形态取决于细胞间信号传导途径和区域特异性转录因子的组合输入。在黑腹果蝇中，蛋白质 Ultrabithorax (Ubx) 是一种区域特异性转录因子，也是同源结构域蛋白 Hox 家族的成员，它通过调节促有丝分裂十触瘫 (Dpp) 途径的几个组件的表达来部分限制后翅 (haltere) 组织的生长。该项目的目标是鉴定果蝇基因组中由haltere中的Ubx直接调控的靶基因和转录调控元件；将使用三种实验方法来实现这一目标。首先，将使用一种新的基因表达谱分析方法来分析连续同源的Ubx表达（haltere）和非表达（wing）组织中的区域基因表达差异。第二步将使用染色质免疫沉淀和 DamID 结合果蝇基因组平铺阵列来确定 haltere 中 Ubx 直接结合的 DNA 区域。总之，这些实验将允许在全基因组范围内鉴定 Haltere 中的直接 Ubx 靶基因。该分析的第三步是使用同源位点识别 (CSI) 微阵列技术来表征 Ubx 的 DNA 结合特异性，无论是作为单体还是与 Hox 辅因子 Extradenticle (Exd) 和 Homothorax (Hth) 结合。这将允许探索 Ubx 可以结合和不能结合的 DNA 序列，并且与前两个步骤相结合，应该可以更好地理解 Hox 调控网络。对 Ubx 靶基因子集的后续分析，特别是那些可能参与生长调节的基因，将包括验证其在翅膀和哈特雷中的表达模式，以及使用标准果蝇遗传方法进行功能分析。公共健康相关性：Hox 蛋白和许多控制生长、增殖和形态发生的信号分子在果蝇和人类之间是保守的；这项研究的目的是深入了解 Hox 转录因子调节组织生长的机制。最终，详细描述 Hox 蛋白在正常发育过程中如何调节组织生长对于理解它们在癌症等疾病状态下生长失调时的作用至关重要。