The functional relationship between the chromatin remodeler dMi-2 and RNA

染色质重塑剂 dMi-2 与 RNA 之间的功能关系

• 批准号: 533518636
• 负责人: Professor Dr. Alexander Brehm
• 金额: --
• 依托单位: Institut für Molekularbiologie und Tumorforschung (IMT)
• 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/533518636?language=en
• 关键词: functional; relationship; between; chromatin; remodeler

Gene expression in eukaryotic cells involves the generation of a conducive chromatin structure, transcription of the DNA template into RNA, co-transcriptional RNA processing, transport into the cytoplasm and translation of the mRNA into protein. Modulation of chromatin structure on the one hand and RNA production and processing on the other have long been viewed as largely separate steps of gene expression. However, recent research suggests the existence of many functional connections but the molecular principles of this interplay are just beginning to emerge. Drosophila Mi-2 is an ATP-dependent nucleosome remodeler with important functions in gene regulation during differentiation. We have recently demonstrated that dMi-2 associates with mRNA in vivo and that this interaction impacts its chromatin binding and nucleosome remodeling activities. We have now found that dMi-2 modulates alternative mRNA splicing. This identifies an unexpected reciprocal relationship between a classical chromatin remodeller and RNA. In this project we will identify the molecular mechanisms underlying the (1) formation and impact of dMi-2:RNA complexes and (2) the modulation of alternative splicing by dMi-2 by using complementary structural, biochemical, genetic and genomic approaches. In the first part of the project we will define the protein and RNA determinants of dMi-2:RNA complex formation, analyse the contributions of dMi-2-associated and RNA bound proteins and define the impact of dMi-2:RNA complexes on transcriptome and chromatin landscape. In the second part of the project we will define the mechanisms by which dMi-2 modulates alternative splicing by determining the contributions of altered RNA polymerase II elongation rate, splicing-associated chromatin modifications, the interplay with splicing factors and dMi2:RNA binding itself. Given the conservation of dMi-2/CHD4 functions and the important roles that dMi-2/CHD4 plays in development and disease we expect our findings to have far reaching implications.

真核细胞中的基因表达涉及有益染色质结构的生成、DNA模板转录成RNA、共转录RNA加工、转运到细胞质以及mRNA翻译成蛋白质。一方面是染色质结构的调节，另一方面是RNA的产生和加工，长期以来一直被视为基因表达的基本独立步骤。然而，最近的研究表明存在许多功能连接，但这种相互作用的分子原理才刚刚开始显现。果蝇 Mi-2 是一种 ATP 依赖性核小体重塑因子，在分化过程中的基因调控中具有重要功能。我们最近证明 dMi-2 在体内与 mRNA 相关，并且这种相互作用影响其染色质结合和核小体重塑活性。我们现在发现 dMi-2 调节选择性 mRNA 剪接。这确定了经典染色质重塑因子和 RNA 之间存在意想不到的相互关系。在这个项目中，我们将通过使用互补的结构、生化、遗传和基因组方法来确定（1）dMi-2：RNA复合物的形成和影响以及（2）dMi-2对选择性剪接的调节背后的分子机制。在该项目的第一部分，我们将定义 dMi-2:RNA 复合物形成的蛋白质和 RNA 决定因素，分析 dMi-2 相关蛋白和 RNA 结合蛋白的贡献，并定义 dMi-2:RNA 复合物对转录组和染色质景观的影响。在该项目的第二部分中，我们将通过确定改变的 RNA 聚合酶 II 延伸率、剪接相关染色质修饰、剪接因子的相互作用以及 dMi2:RNA 结合本身的贡献来定义 dMi-2 调节选择性剪接的机制。鉴于 dMi-2/CHD4 功能的保守性以及 dMi-2/CHD4 在发育和疾病中发挥的重要作用，我们预计我们的发现将产生深远的影响。