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共阻遏物。我们的总体目标是了解细胞信号通路如何在发育过程中介导精确的转录基因控制。我们专注于Notch信号通路及其在指导神经细胞命运和状态中的作用。我们致力于阐明基因调控的一般原则，结合果蝇和哺乳动物模型系统的研究。 该提案描述了第一次在体内的BEN-独奏域蛋白质的功能研究。从我们的果蝇工作中，我们发现BEN-单独因子不敏感（Insv）。通过作为Notch转录因子CSL的直接辅阻遏物调节Notch信号传导和外周神经系统发育。我们将进一步研究其遗传要求，剖析其功能域，并阐明它如何与其他染色质因子合作，诱导转录抑制。我们基于这一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，并且我们有关于它们抑制Notch信号传导并影响神经干细胞自我更新和分化的能力的初步数据。 这些表型驱动的研究将通过对果蝇和小鼠中CSL和BEN-单独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调节的增强子。我们将在体内验证增强子，重点是神经调控元件。我们希望确定神经系统中N/CSL/BEN靶网络的保守特征。此外，这些数据可能揭示了BEN-solo蛋白与染色质的非CSL依赖性关联，我们将使用功能测定来解决这一问题。总的来说，该建议结合了各种实验方法和模型系统，以研究Notch通路中的一种新的保守神经辅阻遏物。

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.