Structure and dynamics of transcriptionally active chromatin domains of Drosophila melanogaster interphase chromosomes

果蝇间期染色体转录活性染色质域的结构和动态

• 批准号: 262515404
• 负责人: Professor Dr. Harald Saumweber
• 金额: --
• 依托单位: Arbeitsgruppe Zytogenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/262515404?language=en
• 关键词: Structure; dynamics; transcriptionally; active; chromatin

Our current data show that the 61C7-8 locus in salivary glands is formed by a ~20 kb open chromatin domain that contains four protein coding genes and one noncoding transcript. By site directed recombination we generated a wild type and partial deleted versions of the locus at the same ectopic position that were initially characterized by cytogenetic methods. In the proposed project we want to study the function of key DNA-elements for the formation of 61C7-8 chromatin at high resolution with the following objectives:1. Determination of the 61C7-8 open domain structure (histone modification and insulator protein binding) in different tissues and stages of development. 2. Determination of the influence of DNA elements on a 61C7-8 domain structure and transcription.3. Determination of the influence of BEAF-binding- and promoter-elements on the 3D folding of the 61C7-8 domain.4. Mapping and initial characterization of BEAF regulated active chromosomal domains in Drosophila.Taking 61C7-8 as a model the experiments will provide understanding on how an open chromatin domain is formed in development. It will further provide information on the stability of basic domain structure and the extent of its modifications in different tissues in correlation to gene expression. The experiments will show the importance of the insulator proteins BEAF, CP190 and Chriz for the formation of chromatin modifications and folding of an open chromatin domain. Moreover, they will address how Chriz and CP190 may contribute to both aspects independently of BEAF binding and will address the mutual dependency of factor binding and transcriptional activity. The elimination of single BEAF binding sites within the domain will allow us to understand the mutual interaction between factors bound at individual sites and possibly the hierarchy of interactions within the domain.Finally, the experiments are intended to extend our 61C7-8 related studies to further open chromatin domains with a similar BEAF connected architecture. Mapping of more open domains will influence our current concepts on polytene and diploid chromatin domain structure, in particular our concepts on interbands as open chromatin domains. In the future the methods established for the 61C7-8 region may be used and extended for the study of open chromatin domains that depend on other insulator proteins.

我们目前的数据表明，61 C7 -8位点在唾液腺中是由一个约20 kb的开放染色质结构域，其中包含四个蛋白质编码基因和一个非编码转录本。通过定点重组，我们产生了一个野生型和部分删除版本的基因座在同一异位位置，最初的特点是细胞遗传学方法。在本项目中，我们希望以高分辨率研究61 C7 -8染色质形成的关键DNA元件的功能，其目标如下：1。不同组织和发育阶段中61 C7 -8开放结构域结构（组蛋白修饰和绝缘子蛋白结合）的测定。2.确定DNA元件对61 C7 -8结构域结构和转录的影响.确定BEAF结合元件和启动子元件对61 C7 -8结构域的3D折叠的影响。果蝇BEAF调控的染色体活性结构域的定位和初步鉴定以61 C7 -8为模型，本实验将为理解一个开放的染色质结构域是如何在发育过程中形成提供帮助。它将进一步提供有关基本结构域结构的稳定性及其在不同组织中与基因表达相关的修饰程度的信息。实验将显示绝缘子蛋白BEAF、CP 190和Chriz对于形成染色质修饰和开放染色质结构域折叠的重要性。此外，他们将解决Chriz和CP 190如何独立于BEAF结合对这两个方面做出贡献，并将解决因子结合和转录活性的相互依赖性。消除结构域中的单个BEAF结合位点将使我们能够理解结合在单个位点上的因子之间的相互作用，以及结构域中相互作用的可能层次。最后，实验旨在扩展我们的61 C7 -8相关研究，以进一步开放具有类似BEAF连接结构的染色质结构域。更多的开放域的映射将影响我们目前的概念，多线和二倍体染色质结构域的结构，特别是我们的概念，带间作为开放的染色质结构域。在未来的61 C7 -8区域建立的方法可以使用和扩展依赖于其他绝缘子蛋白的开放染色质结构域的研究。