ROG--A NOVEL PROTEIN THAT REGULATES TH2 CYTOKINES

描述(改编自《调查者摘要》)：辅助者T的调节失调 (TH)细胞免疫反应是一种主要的致病机制 自身免疫性和过敏性疾病。GATA-3，一种锌指DNA结合蛋白， 在调节细胞的发育、分化和分化方面起着至关重要的作用 Th细胞的活化。特别是，GATA-3仅在类型2中表达 (Th2)似乎调节Th2细胞因子产生的亚群。 因此，改变GATA-3的功能是操纵Th的一种新方法 免疫反应。此前的研究表明，其他GATA的功能 家族成员可以通过与其他蛋白质的相互作用来调节。 最近，我们分离出了一种特定的GATA-3相互作用蛋白，我们称之为 在酵母中通过相互作用陷阱克隆了GATA-3的阻遏因子Rog。罗格是一颗锌 含有“宽阔的复合体、有轨电车轨道和支撑物”的指状蛋白 (Btb)结构域在其N-末端区域。因此，ROG属于一个不断扩大的家庭 含有BTB的锌指蛋白，其中包括转录因子 BCL-6，最近被证明在调节Th免疫反应中起关键作用。罗格 是1型(Th1)和Th2细胞中的早期反应基因，并有效地 抑制白介素4和白介素5的活性 (IL-5)在体外以GATA-3特异性和GATA-3依赖的方式启动。 这些发现提出了许多问题，这是本方案中的实验所针对的 致信地址。首先，ROG是通过什么机制来抑制 GATA-3？ROG可能阻止GATA-3与DNA结合，或者可能形成一个失活的 或与GATA-3的抑制复合体(目标1)。第二，ROG的表达不是 仅限于Th2细胞，提示ROG具有独立于 GATA-3。组织分布和表达动力学的详细分析 Rog，ROG在成熟Th细胞中的过表达，以及ROG的生成和 ROG缺陷小鼠的特征应该提供重要的见解 Rog函数(目标2)。例如，ROG可能会负向调节Th1细胞因子 或其他T细胞特异性基因通过与Th1或T细胞特异性基因相互作用 转录因子。AIM中建议的ROG相互作用蛋白的分离 3可能识别出这些新的组织特异性因子。最后，出现了 多种BTB蛋白，如ROG和Bcl-6，能够抑制Th2 细胞因子基因，提示ROG可能与其他BTB蛋白协同作用 通过抑制作用调节Th细胞的分化和激活 机制(目标4)。

DESCRIPTION (Adapted from Investigator's Abstract): Dysregulation of helper T (Th) cells immune response is a major pathogenic mechanism mediating several autoimmune and allergic diseases. GATA-3, a zinc finger DNA binding protein, plays an essential role in regulating the development, differentiation, and activation of Th cells. In particular, GATA-3 is expressed only in the Type 2 (Th2) subset where it appears to regulate the production of Th2 cytokines. Thus, altering the function of GATA-3 is a novel approach to manipulate the Th immune response. Previous studies have shown that the function of other GATA family members can be modulated by their interaction with other proteins. Recently, we have isolated a specific GATA-3 interacting protein, that we call ROG (Repressor Of GATA-3), by interaction trap cloning in yeast. ROG is a zinc finger protein that contains a "Broad complex, tramtrack, and bric a brac" (BTB) domain in its N-terminal region. Thus, ROG falls into an expanding family of BTB-containing zinc finger proteins which includes the transcription factor Bcl-6, recently shown to be critical in regulating the Th immune response. ROG is an early response gene in both Type 1 (Th1) and Th2 cells and efficiently represses the activity of both the Interleukin-4 (IL-4) and Interleukin-5 (IL-5) promoters in a GATA-3 specific and GATA-3 dependent fashion in vitro. These findings raise many questions which the experiments in this proposal aim to address. First, what is the mechanism by which ROG represses the function of GATA-3? ROG might prevent GATA-3 from binding to DNA or could form an inactive or inhibitory complex with GATA-3 (Aim 1). Second, the expression of ROG is not limited to Th2 cells suggesting that ROG has functions that are independent of GATA-3. Detailed analysis of the tissue distribution and expression kinetics of ROG, overexpression of ROG in mature Th cells, as well as the generation and characterization of ROG deficient mice should provide important insights into ROG function (Aim 2). For example, ROG might negatively regulate Th1 cytokines or other T cell-specific genes by interacting with a Th1- or T cell-specific transcription factor. Isolation of ROG-interacting proteins as proposed in Aim 3 might identify such novel tissue-specific factors. Finally, the presence of multiple BTB proteins, such as ROG and Bcl-6, capable of repressing Th2 cytokine genes, suggests that ROG might act in concert with other BTB proteins to regulate the differentiation and activation of Th cells by inhibitory mechanisms (Aim 4).