REGULATION OF ANDROGEN RECEPTOR ACTIVITY IN THE PROSTATE

我们广泛的、长期的目标是确定雄激素受体(AR)通过其N-末端调节转录激活的分子机制。AR是一种激素依赖的转录因子，参与调节正常和恶性前列腺细胞的生长。虽然雄激素，如双氢睾酮(DHT)是激活AR转录调节功能的主要信号，但AR介导的转录活性也受参与转录的未知调节辅因子的控制。AR的N-末端包含几个转录激活功能，包括AF-1a和AF-1b，这些区域的突变降低了AR依赖的转录活性。我们认为这些结构域提供了允许AR N末端与参与转录调控的辅助因子之间的蛋白质-蛋白质接触的表面。我们认为这些结构域提供了允许AR N末端与参与转录调控的辅助因子之间的蛋白质-蛋白质接触的表面。我们将使用一种改进的酵母双杂交方法来识别与AR N末端激活域相互作用的蛋白质。分离出与转录激活剂相互作用的蛋白质。我们还将定义这些AR与转录激活剂相互作用的效果。我们还将使用瞬时转染系统来确定这些AR相互作用蛋白对AR转录活性的影响，该系统旨在监测培养的哺乳动物细胞中AR依赖的转录激活。可想而知，这些AR N末端相互作用蛋白水平的改变调节AR的活性，因此，如果在癌症中改变，则有助于AR依赖的恶性前列腺生长。为了验证这一假设，我们将使用抗体和核酸探针来确定特定AR相互作用蛋白的浓度在良性和恶性前列腺生长模型中是否有所不同。了解AR和AR N-末端转录激活域相关蛋白之间的通讯是了解AR调控基因表达机制的基础，并可能为开发AR依赖的恶性肿瘤(如前列腺癌)的新疗法提供新的干预点。

DESCRIPTION (Adapted from the application) Our broad, long term objective is to characterize the molecular mechanisms by which the androgen receptor (AR) regulates transcriptional activation through its N-terminus. The AR is a hormone- dependent transcription factor involved in the regulation of both normal and malignant prostate cell growth. Although androgen hormones, such as dihydrotestosterone (DHT), act as the primary signal in activating AR's transcriptional regulatory functions, AR-mediated transcriptional activity is also controlled by associations with, as yet, unidentified regulatory cofactors involved in transcription. The N-terminus of AR contain several transcriptional activation functions, including AF-1a and AF-1b, and mutations in these domains reduce AR-dependent transcriptional activity. We propose that these domains provide surfaces that permit protein-protein contacts between the AR N-terminus and cofactors involved in transcriptional regulation. We propose that these domains provide surfaces that permit protein-protein contacts between the AR N-terminus and cofactors involved in transcriptional regulation. We will identify proteins that interact with the AR N-terminal activation domains using a modified yeast two-hybrid approach that is capable. Of isolating proteins that interact with transcriptional activators.. We will also define the effect of these AR-interact with transcriptional activators. We will also define the effect of these AR-interacting proteins on AR- transcriptional activity using a transient transfection proteins on AR- transcriptional activity using a transient transfection system designed to monitor AR-dependent transcriptional activation in cultured mammalian cells. Conceivably, alterations in the level of these AR N-terminal interacting proteins modulate AR activity., thereby, contributing to malignant AR-dependent prostate growth if altered in cancer. To test this hypothesis we will determine if the concentration of specific AR- interacting proteins varies in models of benign and malignant prostate growth using antibody and nucleic acid probes. Understanding of the communication between AR and the proteins that associate with the AR N-terminal transcriptional activation domain is fundamental to understanding the mechanism of AR-regulated gene expression and may reveal novel points of intervention to be exploited in the development of new therapies for AR-dependent malignancies, such as prostate cancer.