The role of roX RNA for structure and function of the dosage compensation complex.

roX RNA 对剂量补偿复合物结构和功能的作用。

• 批准号: 417339159
• 负责人: Professor Dr. Peter Burkhard Becker
• 金额: --
• 依托单位: Lehrstuhl Molekularbiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/417339159?language=en
• 关键词: role; roX; RNA; structure; function

Complex eukaryotic genomes encode abundant long, non-coding (lnc) RNAs. Our knowledge of the molecular mechanisms underlying chromatin-based lncRNA function is scarce.We will study a paradigmatic, multi-enzyme chromatin regulator, which absolutely requires lncRNA for assembly and function. The dosage compensation complex (DCC) of Drosophila melanogaster consists of five male-specific-lethal (MSL) proteins that assemble with non-coding roX RNA to form a regulatory complex with special properties: the DCC doubles the transcription of most genes on the single X chromosome in male cells to match the combined output of the two Xs in females.Although the protein subunits alone can associate with each other and are active enzymes in vitro, the association of roX is a vital requirement for DCC function in cells. RoX function, however, is mysterious. Secondary structure remodeling of roX by the DExH helicase MLE initiates association of MSL proteins towards assembly of a functional DCC. We hypothesize that the MSL proteins acquire their dedicated functionality within the DCC by association with roX. We aim to elucidate the molecular basis of lncRNA roX integration into the DCC to understand the roX dependency of DCC assembly and activity. To this end, we will study the mechanisms of roX incorporation in detail, optimise the assembly of DCC containing specific RNA and characterize the roX-MSL interactions through specific protein-RNA interaction assays (involving crosslinking, sequencing and mass spectrometry). Our final goal is to compare MSL assemblies in the presence and absence of specific RNA and to correlate structural changes with refined MSL functions.Our study will shed light on a fascinating, highly evolved chromatin regulator. More importantly, we strive to uncover regulatory principles of lncRNA function of general relevance and broad applicability for the emerging field of lncRNA biology.

复杂的真核基因组编码丰富的长非编码（lnc）RNA。我们对基于染色质的lncRNA功能的分子机制的知识是稀缺的。我们将研究一个典型的，多酶染色质调节剂，它绝对需要lncRNA的组装和功能。果蝇的剂量补偿复合物（DCC）由五种雄性特异性致死（MSL）蛋白组成，它们与非编码roX RNA组装形成具有特殊性质的调节复合物：DCC使雄性细胞中单个X染色体上的大多数基因的转录加倍，以匹配雌性细胞中两个X的组合输出。在体外，roX的结合是细胞中DCC功能的重要要求。然而，RoX功能是神秘的。通过DExH解旋酶MLE对roX的二级结构重塑启动MSL蛋白向功能性DCC的组装的缔合。我们假设MSL蛋白通过与roX结合在DCC中获得其专用功能。我们的目的是阐明lncRNA roX整合到DCC的分子基础，以了解DCC组装和活性的roX依赖性。为此，我们将详细研究roX掺入的机制，优化含有特定RNA的DCC的组装，并通过特定的蛋白质-RNA相互作用测定（包括交联，测序和质谱）表征roX-MSL相互作用。我们的最终目标是比较存在和不存在特定RNA的情况下的MSL组装，并将结构变化与精细的MSL功能联系起来。我们的研究将揭示一种迷人的、高度进化的染色质调节因子。更重要的是，我们努力揭示lncRNA生物学新兴领域的普遍相关性和广泛适用性的lncRNA功能的调控原则。