Froesch B A, Takayama S, Reed J C
Burnham Institute, La Jolla, California 92037, USA.
J Biol Chem. 1998 May 8;273(19):11660-6. doi: 10.1074/jbc.273.19.11660.
BAG-1 is a regulator of heat shock protein (Hsp) 70/Hsc70 family proteins that interacts with steroid hormone receptors. The recently identified BAG-1 long (BAG-1L) protein, an isoform of BAG-1 that arises from translation initiation at a noncanonical CUG codon, was co-immunoprecipitated with androgen receptors (AR) from LNCaP prostate cancer cells and other cell lysates, whereas the shorter originally identified BAG-1 and BAG-1M (RAP 46) proteins were not. BAG-1L, but not BAG-1 or BAG-1M (RAP46), also markedly enhanced the ability of AR to transactivate reporter gene plasmids containing an androgen response element (ARE) in PC3 prostate cancer and other cell lines. A C-terminal region deletion mutant of BAG-1L failed to co-immunoprecipitate with AR and functioned as a trans-dominant inhibitor of BAG-1L, impairing AR-induced transactivation of ARE-containing reporter plasmids. In addition, BAG-1L significantly reduced the concentrations of 5alpha-dihydrotestosterone (DHT) required for AR activity but did not induce ligand-independent transactivation. BAG-1L also markedly improved the ability of AR to transactivate reporter genes when cells were cultured with DHT in combination with the anti-androgen cyproterone acetate. The effects of BAG-1L on AR could not be explained by detectable alterations in the DHT-induced translocation of AR from cytosol to nucleus, nor by BAG-1L-induced increases in the amounts of AR protein. These findings implicate BAG-1L in the regulation of AR function and may have relevance to mechanisms of prostate cancer resistance to hormone-ablative and anti-androgen therapy.
BAG-1是一种热休克蛋白(Hsp)70/Hsc70家族蛋白的调节因子,可与类固醇激素受体相互作用。最近鉴定出的BAG-1长链(BAG-1L)蛋白是BAG-1的一种异构体,它由非经典CUG密码子处的翻译起始产生,可与LNCaP前列腺癌细胞和其他细胞裂解物中的雄激素受体(AR)进行共免疫沉淀,而最初鉴定出的较短的BAG-1和BAG-1M(RAP 46)蛋白则不能。BAG-1L而非BAG-1或BAG-1M(RAP46)也显著增强了AR在PC3前列腺癌和其他细胞系中反式激活含有雄激素反应元件(ARE)的报告基因质粒的能力。BAG-1L的C末端区域缺失突变体无法与AR进行共免疫沉淀,并作为BAG-1L的反式显性抑制剂发挥作用,损害AR诱导的含ARE报告质粒的反式激活。此外,BAG-1L显著降低了AR活性所需的5α-二氢睾酮(DHT)浓度,但未诱导非配体依赖性反式激活。当细胞与DHT联合抗雄激素醋酸环丙孕酮一起培养时,BAG-1L也显著提高了AR反式激活报告基因的能力。BAG-1L对AR的作用无法通过DHT诱导的AR从细胞质向细胞核的转位的可检测变化来解释,也无法通过BAG-1L诱导的AR蛋白量的增加来解释。这些发现表明BAG-1L参与AR功能的调节,可能与前列腺癌对激素消融和抗雄激素治疗的耐药机制有关。