REGULATION OF ANDROGEN RECEPTOR ACTIVITY IN THE PROSTATE

我们广泛而长期的目标是描述雄激素受体（AR）通过其n端调节转录激活的分子机制。AR是一种激素依赖性转录因子，参与正常和恶性前列腺细胞生长的调节。虽然双氢睾酮（DHT）等雄激素是激活AR转录调节功能的主要信号，但AR介导的转录活性也受到与转录相关的尚未确定的调节辅助因子的关联控制。AR的n端包含几个转录激活功能，包括AF-1a和AF-1b，这些结构域的突变会降低AR依赖的转录活性。我们认为这些结构域提供了允许AR n端和参与转录调控的辅因子之间的蛋白-蛋白接触的表面。我们认为这些结构域提供了允许AR n端和参与转录调控的辅因子之间的蛋白-蛋白接触的表面。我们将使用一种改良的酵母双杂交方法鉴定与AR n端激活域相互作用的蛋白质。分离与转录激活因子相互作用的蛋白质。我们还将定义这些ar相互作用与转录激活剂的作用。我们还将使用瞬时转染系统来定义这些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。可以想象，这些AR n端相互作用蛋白水平的改变可以调节AR活性。因此，如果在癌症中发生改变，则有助于恶性ar依赖性前列腺生长。为了验证这一假设，我们将使用抗体和核酸探针确定在良性和恶性前列腺生长模型中特异性AR相互作用蛋白的浓度是否不同。了解AR与AR n端转录激活域相关蛋白之间的通讯是理解AR调控基因表达机制的基础，并可能揭示AR依赖性恶性肿瘤（如前列腺癌）新疗法开发的新干预点。

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.