Satoh Tetsurou, Ishizuka Takahiro, Tomaru Takuya, Yoshino Satoshi, Nakajima Yasuyo, Hashimoto Koshi, Shibusawa Nobuyuki, Monden Tsuyoshi, Yamada Masanobu, Mori Masatomo
Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, 3-39-15 Showa-machi, Maebashi, Gunma 371-8511, Japan.
Endocrinology. 2009 Jul;150(7):3283-90. doi: 10.1210/en.2008-1122. Epub 2009 Mar 26.
The 26S proteasome, which degrades ubiquitinated proteins, appears to contribute to the cyclical loading of androgen receptor (AR) to androgen response elements of target gene promoters; however, the mechanism whereby the 26S proteasome modulates AR recruitment remains unknown. Using yeast two-hybrid screening, we previously identified Tat-binding protein-1 (TBP-1), an adenosine triphosphatase of 19S regulatory particles of the 26S proteasome, as a transcriptional coactivator of thyroid hormone receptor. Independently, TBP-1-interacting protein (TBPIP) was also identified as a coactivator of several nuclear receptors, including AR. Here, we investigated whether TBP-1 could interact with and modulate transcriptional activation by AR cooperatively with TBPIP. TBP-1 mRNA was ubiquitously expressed in human tissues, including the testis and prostate, as well as in LNCaP cells. TBP-1 directly bound TBPIP through the amino-terminal domain possessing the leucine zipper structure. AR is physically associated with TBP-1 and TBPIP in vitro and in LNCaP cells. TBP-1 similarly and additively augmented AR-mediated transcription upon coexpression with TBPIP, and the ATPase domain, as well as leucine zipper structure in TBP-1, was essential for transcriptional enhancement. Overexpression of TBP-1 did not alter AR protein and mRNA levels. In the chromatin immunoprecipitation assay, TBP-1 was transiently recruited to the proximal androgen response element of the prostate-specific antigen gene promoter in a ligand-dependent manner in LNCaP cells. These findings suggest that a component of 19S regulatory particles directly binds AR and might participate in AR-mediated transcriptional activation in cooperation with TBPIP.
26S蛋白酶体可降解泛素化蛋白,它似乎有助于雄激素受体(AR)周期性地加载到靶基因启动子的雄激素反应元件上;然而,26S蛋白酶体调节AR募集的机制尚不清楚。通过酵母双杂交筛选,我们之前鉴定出Tat结合蛋白1(TBP-1),它是26S蛋白酶体19S调节颗粒的一种三磷酸腺苷酶,作为甲状腺激素受体的转录共激活因子。另外,TBP-1相互作用蛋白(TBPIP)也被鉴定为包括AR在内的几种核受体的共激活因子。在此,我们研究了TBP-1是否能与TBPIP协同作用,与AR相互作用并调节其转录激活。TBP-1 mRNA在包括睾丸和前列腺在内的人体组织以及LNCaP细胞中普遍表达。TBP-1通过具有亮氨酸拉链结构的氨基末端结构域直接与TBPIP结合。在体外和LNCaP细胞中,AR与TBP-1和TBPIP存在物理关联。与TBPIP共表达时,TBP-1同样且额外地增强了AR介导的转录,并且TBP-1中的ATP酶结构域以及亮氨酸拉链结构对于转录增强至关重要。TBP-1的过表达并未改变AR蛋白和mRNA水平。在染色质免疫沉淀试验中,在LNCaP细胞中,TBP-1以配体依赖方式被短暂募集到前列腺特异性抗原基因启动子的近端雄激素反应元件上。这些发现表明,19S调节颗粒的一个组分直接结合AR,并可能与TBPIP协同参与AR介导的转录激活。