BEN factors are conserved CSL co-repressors in Notch-mediated neural development

描述（由申请人提供）：BEN因子是notch介导的神经发育中保守的CSL共抑制因子。我们的总体目标是了解细胞信号通路在发育过程中如何介导准确的转录基因控制。我们关注Notch信号通路及其在指导神经细胞命运和状态中的作用。我们努力通过结合果蝇和哺乳动物模型系统的研究来阐明基因调控的一般原理。本研究首次描述了BEN-solo结构域蛋白的体内功能研究。从我们对果蝇的研究中，我们发现BEN-solo因子不敏感（Insv）。作为Notch转录因子CSL的直接共抑制因子，调节Notch信号和周围神经系统的发育。我们将进一步研究其遗传需求，解剖其功能域，并阐明其如何与其他染色质因子合作诱导转录抑制。我们利用这些专有知识来研究哺乳动物BEN-solo因子在神经发育过程中的作用，并且我们有初步数据表明它们能够抑制Notch信号传导并影响神经干细胞的自我更新和分化。这些表型驱动的研究将通过对果蝇和小鼠中CSL和BEN-solo因子的染色质结合的全基因组分析来补充，我们将从中提取notch调节增强子。我们将在体内验证增强剂，重点是神经调节元件。我们希望确定N/CSL/BEN靶网络在神经系统中的保守特征。此外，这些数据可能揭示了与csl无关的BEN-solo蛋白与染色质的关联，我们将使用功能分析来解决这个问题。总的来说，本研究结合了多种实验方法和模型系统来研究Notch通路中一种新的保守神经共抑制因子。

DESCRIPTION (provided by applicant): BEN factors are conserved CSL co-repressors in Notch-mediated neural development. Our general goal is to understand how cell signaling pathways mediate accurate transcriptional gene control during development. We focus on the Notch signaling pathway and its roles in directing neural cell fates and states. We strive to elucidate general principles of gene regulation by combining studies in both Drosophila and mammalian model systems. This proposal describes first in vivo functional studies of BEN-solo domain proteins. From our Drosophila work, we found that the BEN-solo factor Insensitive (Insv). regulates Notch signaling and peripheral nervous system development by acting as a direct corepressor for the Notch transcription factor CSL. We will further study its genetic requirements, dissect its functional domains, and elucidate how it cooperates with other chromatin factors to induce transcriptional repression. We build upon this proprietary knowledge to study roles of mammalian BEN-solo factors during neural development, and we have preliminary data on their ability to inhibit Notch signaling and affect neural stem cell self-renewal and differentiation. These phenotype-driven studies will be complemented by genomewide analyses of chromatin binding of CSL and BEN-solo factors in flies and mice, from which we will extract Notch-regulated enhancers. We will validate enhancers in vivo with emphasis on neural regulatory elements. We hope to determine conserved features of N/CSL/BEN target networks in the nervous system. In addition, these data potentially shed light on CSL-independent association of BEN-solo proteins with chromatin, which we will address using functional assays. Overall, this proposal combines a variety of experimental approaches and model systems to investigate a novel conserved neural corepressor in the Notch pathway.