Re-evaluating enhancer modularity and boundaries with quantitative approaches in Drosophila

用果蝇的定量方法重新评估增强子的模块化和边界

• 批准号: 531924191
• 负责人: Professor Dr. Nicolas Gompel
• 金额: --
• 依托单位: Abteilung III: Evolutionsbiologie und Ökologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/531924191?language=en
• 关键词: Re; evaluating; enhancer; modularity; boundaries

Transcriptional enhancers control in which cells, at what time, and how much a given gene is transcribed from a core promoter. The precision of this process is for instanceb necessary for normal embryonic development of all multicellular organisms. Classically, enhancers have been revealed and defined in reporter assays, where the regulatory activity of a DNA segment is tested in cell culture, or in transgenic organisms. This approach has painted a textbook picture where enhancers are short (100-1000 bp), discrete stretches of DNA sequence, working as independent modules. Using a different and quantitative approach to map enhancers, we have recently found that enhancers of the pigmentation gene yellow, driving activity in the wings of flies (Drosophila) appear much broader (~4000 bp) and entangled than the literature predicts. Preliminary evidence indicates that the results extend to another enhancer of yellow driving in the fly body. This situation undermines the notion of enhancer modularity, a key concept to understand Developmental and Evolutionary Biology. With this proposal, we would like to first thoroughly describe the degree to which regulatory information is intermingled among these different enhancers. We then want to understand through which mechanisms enhancers achieve their functional modularity. We suspect that regulatory information may be selectively available in different tissues by means of regulated accessibility of the DNA. We will specifically test this hypothesis by investingating the determinants of enhancer accessibility, in cis and in trans, for the body and wing enhancers. We will combine reporter assays in transgenic Drosophila, computational and quantitative image analysis, and genomic methods to achieve these goals. We expect our results to seed the redefinition of transcriptional enhancers, but also to reveal how regulatory information from different, overlapping enhancers is mobilized in a selective manner.

转录增强子控制在哪些细胞中，在什么时间，以及给定基因从核心启动子转录多少。例如，这个过程的精确性对于所有多细胞生物的正常胚胎发育是必要的。传统上，增强子已经在报告基因测定中被揭示和定义，其中DNA片段的调节活性在细胞培养物或转基因生物中被测试。这种方法描绘了一幅教科书般的画面，其中增强子很短（100-1000 bp），是DNA序列的离散片段，作为独立的模块工作。使用一种不同的和定量的方法来映射增强子，我们最近发现，色素沉着基因的增强子，在苍蝇（果蝇）的翅膀驱动活动似乎更广泛（~4000 bp）和纠缠比文献预测。初步证据表明，结果扩展到另一个增强剂的黄色驾驶在苍蝇的身体。这种情况破坏了增强子模块化的概念，这是理解发育和进化生物学的关键概念。有了这个建议，我们想首先彻底描述的程度，其中监管信息是混杂在这些不同的增强子。然后，我们想了解增强子通过哪些机制实现其功能模块化。我们怀疑，调节信息可能是有选择地在不同的组织中通过调节DNA的可及性。我们将通过研究增强子可及性的决定因素来具体检验这一假设，包括体增强子和翼增强子的顺式和反式。我们将结合联合收割机的报告基因分析，在转基因果蝇，计算和定量图像分析，和基因组的方法来实现这些目标。我们希望我们的研究结果能够为转录增强子的重新定义提供种子，同时也揭示了来自不同的重叠增强子的调控信息是如何以选择性的方式被调动的。