Schiffer Niclas W, Céraline Jocelyn, Hartl F Ulrich, Broadley Sarah A
Department of Cellular Biochemistry, Max Planck Institute of Biochemistry, D-82152 Martinsried, Germany.
Biol Chem. 2008 Dec;389(12):1455-66. doi: 10.1515/BC.2008.169.
Abstract Several neurodegenerative diseases, including Kennedy's disease (KD), are associated with misfolding and aggregation of polyglutamine (polyQ)-expansion proteins. KD is caused by a polyQ-expansion in the androgen receptor (AR), a key player in male sexual differentiation. Interestingly, KD patients often show signs of mild-to-moderate androgen insensitivity syndrome (AIS) resulting from AR dysfunction. Here, we used the yeast Saccharomyces cerevisiae to investigate the molecular mechanism behind AIS in KD. Upon expression in yeast, polyQ-expanded N-terminal fragments of AR lacking the hormone binding domain caused a polyQ length-dependent growth defect. Interestingly, while AR fragments with 67 Q formed large, SDS-resistant inclusions, the most pronounced toxicity was observed upon expression of 102 Q fragments which accumulated exclusively as soluble oligomers in the 100-600 kDa range. Analysis using a hormone-dependent luciferase reporter revealed that full-length polyQ-expanded AR is fully functional in transactivation, but becomes inactivated in the presence of the corresponding polyQ-expanded N-terminal fragment. Furthermore, the greatest impairment of AR activity was observed upon interaction of full-length AR with soluble AR fragments. Taken together, our results suggest that soluble polyQ-containing fragments bind to full-length AR and inactivate it, thus providing insight into the mechanism behind AIS in KD and possibly other polyglutamine diseases, such as Huntington's disease.
摘要 包括肯尼迪病(KD)在内的几种神经退行性疾病与多聚谷氨酰胺(polyQ)扩展蛋白的错误折叠和聚集有关。KD是由雄激素受体(AR)中的多聚谷氨酰胺扩展引起的,AR是男性性别分化中的关键因子。有趣的是,KD患者常表现出由AR功能障碍导致的轻度至中度雄激素不敏感综合征(AIS)的症状。在此,我们利用酿酒酵母来研究KD中AIS背后的分子机制。在酵母中表达时,缺乏激素结合结构域的AR的多聚谷氨酰胺扩展N端片段导致了多聚谷氨酰胺长度依赖性的生长缺陷。有趣的是,虽然含有67个谷氨酰胺的AR片段形成了大的、耐十二烷基硫酸钠的包涵体,但在表达102个谷氨酰胺的片段时观察到最明显的毒性,这些片段仅作为100 - 600 kDa范围内的可溶性寡聚体积累。使用激素依赖性荧光素酶报告基因进行的分析表明,全长多聚谷氨酰胺扩展的AR在反式激活中完全有功能,但在存在相应的多聚谷氨酰胺扩展N端片段时会失活。此外,在全长AR与可溶性AR片段相互作用时观察到AR活性的最大损伤。综上所述,我们的结果表明,含多聚谷氨酰胺的可溶性片段与全长AR结合并使其失活,从而为KD以及可能的其他多聚谷氨酰胺疾病(如亨廷顿病)中AIS背后的机制提供了见解。