KANSAS U COBRE: REGULATION OF GENE EXPRESSION IN THE TH2 CYTOKINE LOCUS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在Th2分化过程中，Th2细胞因子基因座经历了染色质结构的变化，从而促进了细胞因子基因的协调表达。各种积极和消极的调节因素协同工作，以特定血统的方式带来这些变化。这项研究的总体科学目标是了解在该基因座内协调基因表达的顺式元件之间的功能相互作用。我们还想了解转录因子启动和维持细胞因子基因转录活性的机制。以下目标旨在创建一个模型系统，在该模型系统中，可以检查各种顺式元件在Th2细胞因子基因座内转录调控中所起的作用。它们的完成将为成功实现总体科学目标奠定基础。第一个目标是确定在该基因座中发挥结构或功能作用的反式因子结合位点。我们将特别关注两个重要的Th2相关因子GATA3和STAT6的结合位点。将使用生物化学和分子生物学方法来实现这一目标。第二个目标是建立一个报告系统，以评估Th2细胞因子基因座内的元件在调节IL4、IL5和IL13协调表达中的作用。这位记者将被用来同时评估基因的转录，以及评估顺式元件单独和组合在其调控中的作用。适当的Th2分化对于适当的免疫动态平衡和抗原反应性至关重要。调节失调的T细胞极化与许多病理状态有关，包括变态反应性疾病和自身免疫。从这些研究中收集的信息将有助于我们对Th2分化的理解，并有望实现最终目标，干预这一过程，避免或纠正这些病理表现。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. During Th2 differentiation, the Th2 cytokine locus undergoes changes in chromatin structure that facilitate coordinated cytokine gene expression. A variety of positive and negative regulatory elements work cooperatively to bring about these changes in a lineage-specific fashion. The overall scientific goal of this study is to understand the functional interplay among cis-elements that coordinates gene expression within this locus. We also want to understand the mechanism by which transcription factors initiate and maintain transcriptional activity of the cytokine genes. The following aims are designed to create a model system in which the role that various cis-elements play in transcriptional regulation within the Th2 cytokine locus can be examined. Their completion will lay the groundwork for successfully addressing the overall scientific goal. The first aim is to identify trans-factor binding sites serving structural or functional roles in mediating transcriptional activity in the locus. We will specifically focus on binding sites for two important Th2-related factors, GATA3 and STAT6. Biochemical and molecular biological approaches will be used to fulfill the objectives of this aim. The second aim is to generate a reporter system to evaluate the role of elements within the Th2 cytokine locus in regulating coordinated expression of IL4, IL5, and IL13. This reporter will be utilized to assess transcription of the genes simultaneously as well as assess the role of cis-elements, individually and in combination, on their regulation. Appropriate Th2 differentiation is critical for proper immune homeostasis and antigen responsiveness. Dysregulated T cell polarization has been implicated in a number of pathological states, including allergic diseases and autoimmunity. Information gleaned from these studies will contribute to our understanding of Th2 differentiation and hopefully enable the ultimate goal, to intervene in this process and to avoid or correct these pathological manifestations.