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Structure and Function of a Signal-Regulated Developmental Enhancer

信号调节发育增强剂的结构和功能

基本信息

批准号：
8134830
负责人：
Scott Barolo
金额：
$ 28.31万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-14 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8134830
关键词：
Accounting Adult Animals Binding Binding Sites Biochemical Biochemistry Bioinformatics Biological Assay Biological Models Cell Survival Cells Clinical Research DNA DNA Binding DNA Sequence Defect Development Disease Drosophila genus Elements Enhancers Epidermal Growth Factor Receptor Eye Gene Activation Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Genetic Techniques Genetic Transcription Health Imagery In Vitro Introns Investigation Knowledge Link MAP Kinase Gene Malignant Neoplasms Manuscripts Microscopic Molecular Nuclear Nuclear Extract Nucleosomes Organism Pathway interactions Pattern Play Positioning Attribute Protein Family Recording of previous events Regulatory Element Reporter Research Retinal Cone Role Signal Pathway Signal Transduction Site Structure Testing Time Tissues Transcription Coactivator Transcriptional Activation Transcriptional Regulation Transgenic Organisms Work cell motility cell type chromatin modification design developmental genetics disorder prevention functional gain human disease in vivo innovation intercellular communication notch protein novel programs runx proteins tool transcription factor treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Cell-cell signaling pathways, such as Wnt, Notch, and MAPK, are critical for the proper fate specification of most cells and tissues during animal development. These ancient and highly conserved pathways regulate developmental cell fate and adult health primarily by altering the activity of transcription factors (TFs), which in turn control the expression of target genes. Signal-regulated enhancers (or cis-regulatory elements) located near pathway target genes contain binding sites for signal-regulated TFs and for other, tissue- or cell-type- specific TFs; interactions among these factors determine the expression pattern of the target gene. However, no signal-regulated enhancer in any multicellular organism has been fully characterized, in the sense that all of its necessary direct regulatory inputs have been accounted for. Furthermore, at the mechanistic level, none of the known biochemical activities implicated in transcriptional activation have been shown to be sufficient, alone or together, to drive gene expression during animal development. These and other gaps in our understanding suggest that important mechanisms of transcriptional activation are likely still undiscovered. The proposed research program is dedicated to "solving" two signal-regulated developmental enhancers; that is, to defining and functionally characterizing all of their required regulatory sub-elements. Our results, along with other studies, show definitively that simply combining the known regulatory TF sites of well-characterized enhancers is not sufficient to stimulate transcription in vivo. The project proposed here combines in vivo functional reporter assays with the investigation of known transcriptional control mechanisms, the identification of novel regulators, and the advantages of Drosophila genetics. The project has two main objectives: (1) Identify the DNA elements within the sparkling (spa) and dppVM enhancers that are necessary for gene activation in vivo; determine the topological/structural rules for arrangement of those elements. (2) Determine the roles of the known and novel regulatory elements within spa and dppVM in mechanisms of transcriptional control such as local chromatin modification, cooperative DNA binding, nucleosome positioning, and intra-nuclear organization; isolate and characterize factors binding to novel regulatory DNA elements within these enhancers. Project Narrative The signaling pathways and factors to be studied are critical for normal animal development, and are implicated in a host of developmental defects and human diseases, most notably cancer. A better mechanistic understanding of signal-regulated enhancers will facilitate the development of strategies for the treatment and prevention of disorders caused by aberrant cell signaling and gene expression, and may further the development of new molecular tools with significant clinical and research utility.

描述（由申请方提供）：细胞-细胞信号传导途径，如Wnt、Notch和MAPK，对于动物发育期间大多数细胞和组织的正确命运规范至关重要。这些古老且高度保守的途径主要通过改变转录因子（TF）的活性来调节发育细胞命运和成人健康，转录因子反过来控制靶基因的表达。位于通路靶基因附近的信号调节增强子（或顺式调节元件）含有信号调节TF和其他组织或细胞类型特异性TF的结合位点;这些因子之间的相互作用决定靶基因的表达模式。然而，在任何多细胞生物中没有信号调节增强子已被充分表征，在这个意义上，其所有必要的直接调控输入已被占。此外，在机制水平上，没有一种已知的涉及转录激活的生物化学活性被证明是足够的，单独或一起，在动物发育过程中驱动基因表达。这些和我们理解中的其他差距表明，转录激活的重要机制可能仍未被发现。拟议的研究计划是致力于“解决”两个信号调节发育增强剂，也就是说，定义和功能特性的所有所需的监管子元素。我们的研究结果，沿着与其他研究，明确表明，简单地结合已知的调节TF网站的充分表征的增强子是不足以刺激体内转录。在这里提出的项目结合了在体内的功能报告基因检测与已知的转录控制机制的调查，新的监管机构的识别，和果蝇遗传学的优势。该项目有两个主要目标：（1）确定体内基因激活所必需的sparkling（spa）和dppVM增强子内的DNA元件;确定这些元件排列的拓扑/结构规则。(2)确定已知的和新的调控元件内温泉和dppVM的转录控制机制，如局部染色质修饰，合作的DNA结合，核小体定位，和核内组织的作用;分离和表征这些增强子内的新的调控DNA元件结合的因素。项目叙述待研究的信号通路和因子对正常动物发育至关重要，并且与许多发育缺陷和人类疾病（最显著的是癌症）有关。更好地理解信号调节增强子的机制将有助于开发治疗和预防异常细胞信号传导和基因表达引起的疾病的策略，并可能进一步开发具有重要临床和研究效用的新分子工具。