Thomas Monzy, Dadgar Nahid, Aphale Abhishek, Harrell Jennifer M, Kunkel Robin, Pratt William B, Lieberman Andrew P
Department of Pathology, The University of Michigan Medical School, Ann Arbor, Michigan, 48109, USA.
J Biol Chem. 2004 Feb 27;279(9):8389-95. doi: 10.1074/jbc.M311761200. Epub 2003 Dec 11.
Kennedy's disease is a degenerative disorder of motor neurons caused by the expansion of a glutamine tract near the amino terminus of the androgen receptor (AR). Ligand binding to the receptor is associated with several post-translational modifications, but it is poorly understood whether these affect the toxicity of the mutant protein. Our studies now demonstrate that mutation of lysine residues in wild-type AR that are normally acetylated in a ligand-dependent manner mimics the effects of the expanded glutamine tract on receptor trafficking, misfolding, and aggregation. Mutation of lysines 630 or 632 and 633 to alanine markedly delays ligand-dependent nuclear translocation. The K632A/K633A mutant also undergoes ligand-dependent misfolding and aggregation similar to the expanded glutamine tract AR. This acetylation site mutant exhibits ligand-dependent 1C2 immunoreactivity, forms aggregates that co-localize with Hsp40, Hsp70, and the ubiquitin-protein isopeptide ligase (E3) ubiquitin ligase carboxyl terminus of Hsc70-interacting protein (CHIP), and inhibits proteasome function. Ligand-dependent nuclear translocation of the wild-type receptor and misfolding and aggregation of the K632A/K633A mutant are blocked by radicicol, an Hsp90 inhibitor. These data identify a novel role for the acetylation site as a regulator of androgen receptor subcellular distribution and folding and indicate that ligand-dependent aggregation is dependent upon intact Hsp90 function.
肯尼迪病是一种运动神经元退行性疾病,由雄激素受体(AR)氨基末端附近的谷氨酰胺序列扩增引起。配体与受体结合会引发多种翻译后修饰,但这些修饰是否影响突变蛋白的毒性尚不清楚。我们的研究表明,野生型AR中通常以配体依赖方式发生乙酰化的赖氨酸残基发生突变,会模拟扩增的谷氨酰胺序列对受体运输、错误折叠和聚集的影响。将赖氨酸630、632和633突变为丙氨酸会显著延迟配体依赖的核转位。K632A/K633A突变体也会发生与扩增的谷氨酰胺序列AR类似的配体依赖的错误折叠和聚集。这种乙酰化位点突变体表现出配体依赖的1C2免疫反应性,形成与热休克蛋白40(Hsp40)、热休克蛋白70(Hsp70)以及泛素-蛋白异肽连接酶(E3)即热休克蛋白70相互作用蛋白(CHIP)的泛素连接酶羧基末端共定位的聚集体,并抑制蛋白酶体功能。野生型受体的配体依赖核转位以及K632A/K633A突变体的错误折叠和聚集被热休克蛋白90(Hsp90)抑制剂根赤壳菌素阻断。这些数据确定了乙酰化位点作为雄激素受体亚细胞分布和折叠调节因子的新作用,并表明配体依赖的聚集依赖于完整的Hsp90功能。