AUXILIARY PROTEINS IN THE RESPONSE TO THYROID HORMONE

甲状腺激素对糖尿病的正常发育和维持至关重要。 新陈代谢正常。它们影响代谢率，中间代谢， 以及肺部、骨骼系统和中枢的生长和发育 神经系统。甲状腺激素的作用很大程度上是由 与特定节段结合的核甲状腺激素受体(TR) DNA(甲状腺激素反应元件；tre)和调节基因 以一种未知的机制转录。一种核蛋白，称为tr 辅助蛋白(TRAP)，最近被证明可以增强这种能力 结合DNA的受体。TRAP是一种序列特异性DNA结合蛋白 它似乎与tr形成异二聚体。这样做的长期目标是 该项目旨在了解TRAP在TRR功能中的作用。这个 第一个特定目标是确定TRAP所针对的DNA序列的特征 捆绑。用足迹法研究TRAP与DNA的相互作用 分析，以及TRAP DNA结合部位和 TRDNA结合位点将在几个TRE中进行检测。第二 目的是分离和鉴定一个TRAP基因克隆。克隆人 将通过用TRAP筛选1gt11表达文库来分离 DNA结合序列；或通过TRAP的纯化，对 蛋白质，并利用寡核苷酸基于 蛋白质序列。将对cDNA的序列进行检查 与转录因子家族相似。第三个具体目标 目的是研究TRAP在T3应答中的作用。转染性研究 将研究不同的TRE调节A基因表达的能力 报告基因，以及TRA内的点突变的影响 DNA结合部位。此外，TRAP对T3反应的影响将是 通过导入TRAP基因或表达TRAP基因进行研究 反义RNA。第四个具体目标是研究 不同组织中TRAP基因的表达及TRAP家族的可能性 MRNAs。第五个具体目的是检查生理调节。 对TRAP基因表达的影响。所有这些研究的数据都将有所帮助 确定TRAP-TR-DNA复合体的结构，以及TRAP在 T3反应。

Thyroid hormones are essential for normal development and maintenance of normal metabolism. They affect metabolic rate, intermediary metabolism, and growth and development of the lungs, skeletal system, and central nervous system. The effects of thyroid hormone are largely mediated by nuclear thyroid hormone-receptors (TR), which bind to specific segments of DNA (thyroid hormone response elements; TRE) and regulate gene transcription by an unknown mechanism. A nuclear protein, called TR auxiliary protein (TRAP), has recently been shown to enhance the ability of TR to bind to DNA. TRAP is a sequence-specific DNA-binding protein which appears to form heterodimers with TR. The long-term goal of this project is to understand the role of TRAP in the function of TR. The first specific aim is to characterize the DNA sequence to which TRAP binds. The interactions of TRAP with DNA will be studied by footprint analysis, and the relative positions of the TRAP DNA-binding site and the TR DNA-binding site will be examined in several TREs. The second specific aim is to isolate and characterize a TRAP cDNA clone. The clone will be isolated by screening 1gt11 expression libraries with the TRAP DNA-binding sequence; or by purification of TRAP, microsequencing the protein, and screening libraries with oligonucleotides based on the protein sequence. The sequence of the cDNA will be examined for similarity to families of transcription factors. The third specific aim is to study the role of TRAP in the response to T3. Transfection studies will examine the ability of different TREs to regulate expression of a reporter gene, and the effect of point mutations within the TRA DNA-binding site. Also, the effect of TRAP on the response to T3 will be investigated by transfection of the TRAP cDNA, or expression of TRAP antisense RNA. The fourth specific aim is to study the expression of TRAP mRNA in different tissues and the possibility of a family of TRAP mRNAs. The fifth specific aim is to examine the physiological regulation of expression of TRAP mRNA. The data from all these studies will help define the structure of the TRAP-TR-DNA complex, and the role of TRAP in the T3 response.