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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 研究中心，而研究中心不一定是研究者所在的机构。 在Th 2分化过程中，正调控元件和负调控元件在Th 2细胞因子基因座中协同工作，以引起染色质结构的变化，从而以谱系特异性方式促进协调的细胞因子基因表达。本研究的总体目标是了解协调该基因表达的顺式元件之间的相互作用。我们将创建一个模型系统，其中各种顺式元件在Th 2细胞因子基因座内的转录调控中发挥的作用可以进行检查。这些项目的完成将为成功实现总体科学目标奠定基础。 第一个目的是确定在基因座中介导转录活性的结构或功能作用的反式因子结合位点。我们将重点关注两个重要的Th 2相关因子GATA 3和STAT 6的结合位点。生物化学和分子生物学方法将用于实现这一目标。第二个目的是产生报告系统，以评估Th 2细胞因子基因座内的元件在调节IL 4、IL 5和IL 13的协调表达中的作用。该报告基因将用于同时评估基因的转录，以及评估顺式元件单独和组合对其调控的作用。 适当的Th 2分化对于适当的免疫稳态和抗原应答是至关重要的。异常调节的T细胞极化与过敏性疾病和自身免疫有关。从这些研究中收集到的信息将有助于我们理解Th 2分化，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, positive and negative regulatory elements work cooperatively in the Th2 cytokine locus to bring about changes in chromatin structure that facilitate coordinated cytokine gene expression in a lineage-specific fashion. The overall goal of this study is to understand the interplay among cis-elements that coordinates this gene expression. We will 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 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 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.