Whole-genome analysis of direct Doublesex target genes

描述（由申请人提供）：创造不同的雄性和雌性形式（性二态性）是有性生殖动物发育的关键步骤。虽然启动性别决定的机制在不同物种之间有很大差异，但最近的工作表明，性别决定控制创造性二型性的途径更为保守。一个很好的例子是Doublemex Mab 3相关转录因子（DMRT）家族，它已被证明可以调节苍蝇，蠕虫，鱼类，鸟类，小鼠和人类等多种生物体的性二型性。然而，这些关键转录因子控制性二型性的分子途径仍然很大程度上未被探索。 DMRT家族的创始成员是果蝇蛋白Doublemex（DSX），其是控制果蝇中雄性与雌性形态的主要因素。dsx RNA在雄性与雌性中剪接成不同的形式，其编码雄性和雌性蛋白质DSXM和DSXF。这两个高度相关的转录因子如何调节相反的男性与女性发育程序是该领域的一个关键问题，但DSX的靶点很少被确定。我们建议通过采用全基因组分子方法来揭示由DSXM和DSXF控制的调控网络来解决这个问题。我们将使用染色质免疫沉淀，然后进行高通量测序（ChIP-seq），以确定基因组中的DSX结合位点。然后，我们将这些数据与男性与女性的RNA表达谱进行比较，以鉴定DSX靶基因。 这些研究将使我们能够确定DSX控制性腺性二态性的调控网络，并解决有关DSX的男性和女性形式如何差异化调控该网络的基本问题。这也将为未来的工作奠定坚实的基础，重点是DSXM和DSXF用于调节其目标的机制，以及这些目标在控制性二态发育中发挥的作用。这项工作将帮助我们了解人类性发育障碍，如影响DMRTs的疾病，并将提供一个框架，以了解转录因子的选择性剪接一般如何导致下游反应的多样化。 公共卫生相关性：这项工作研究如何在发展过程中调节性别之间的差异。这与我们对人类性发育障碍（DSDs）的理解有关，DSDs会导致性分化和不育的缺陷。

DESCRIPTION (provided by applicant): The creation of distinct male and female forms (sexual dimorphism) is a critical step in the development of sexually reproducing animals. Although the mechanisms that initiate sex determination vary considerably between different species, recent work demonstrates that the pathways that sex determination controls to create sexual dimorphism are more well-conserved. An excellent example is the Doublesex Mab3 Related Transcription Factor (DMRT) family, which have been shown to regulate sexual dimorphism in organisms as diverse as flies, worms, fish, birds, mice and man. However, the molecular pathways by which these key transcription factors control sexual dimorphism remain largely unexplored. The founding member of the DMRT family is the Drosophila protein Doublesex (DSX), which is the main factor controlling male vs. female morphology in flies. The dsx RNA is spliced into distinct forms in males vs. females, which encode the male and female proteins, DSXM and DSXF. How these two, highly-related transcription factors regulate the opposing male vs. female developmental programs is a key question in the field, but few targets for DSX have been identified. We propose to address this question by taking a whole-genome molecular approach to uncover the regulatory network(s) controlled by DSXM and DSXF. We will use chromatin immunoprecipitation followed by high throughput sequencing (ChIP-seq) to identify DSX binding sites in the genome. We will then compare these data to RNA expression profiles on males vs. females to identify DSX target genes. These studies will allow us to identify the regulatory network through which DSX controls gonad sexual dimorphism and address fundamental questions about how the male and female forms of DSX differentially regulate this network. This will also create a strong foundation for future work that focuses on the mechanisms that DSXM and DSXF use to regulate their targets, and what role these targets play in controlling sexually dimorphic development. This work will help us to understand human disorders of sexual development, such as those affecting DMRTs, and will provide a framework for understanding how the alternative splicing of transcription factors in general can lead to a diverse repertoire of downstream responses. PUBLIC HEALTH RELEVANCE: This work studies how the differences between the sexes are regulated during development. This is relevant to our understanding of human Disorders of Sex Development (DSDs), which cause defects in sexual differentiation and infertility.