Welch W J, Diamond M I
Department of Neurology, University of California, San Francisco, CA 94143-0450, USA.
Hum Mol Genet. 2001 Dec 15;10(26):3063-74. doi: 10.1093/hmg/10.26.3063.
Spinobulbar muscular atrophy is a progressive motor neuron disease caused by abnormal polyglutamine tract expansion in the androgen receptor (AR) gene, and is part of a family of central nervous system (CNS) neurodegenerative diseases, including Huntington's disease (HD). Each pathologic protein is widely expressed, but the cause of neuronal degeneration within the CNS remains unknown. Many reports now link abnormal polyglutamine protein aggregation to pathogenesis. A previous study reported that activation of the wild-type glucocorticoid receptor (wtGR) suppressed the aggregation of expanded polyglutamine proteins derived from AR and huntingtin, whereas a mutant receptor containing an internal deletion, GRDelta108-317, increased polyglutamine protein aggregation, in this case primarily within the nucleus. In this study, we use these two forms of GR to study expanded polyglutamine AR protein in different cell contexts. Using cell biology and biochemical approaches, we find that wtGR promotes soluble forms of the protein and prevents nuclear aggregation in NIH3T3 cells and cultured neurons. In contrast, GRDelta108-317 decreases polyglutamine protein solubility, and causes formation of nuclear aggregates in non-neuronal cells. Nuclear aggregates recruit hsp72 more rapidly than cytoplasmic aggregates, and are associated with decreased cell viability. Limited proteolysis and chemical cross-linking suggest unique soluble forms of the expanded AR protein underlie these distinct biological activities. These observations provide an experimental framework to understand why expanded polyglutamine proteins may be toxic only to certain populations of cells, and suggest that unique protein associations or conformations of expanded polyglutamine proteins may determine subsequent cellular effects such as nuclear localization and cellular toxicity.
脊髓延髓肌肉萎缩症是一种由雄激素受体(AR)基因中异常的多聚谷氨酰胺序列扩增引起的进行性运动神经元疾病,它是中枢神经系统(CNS)神经退行性疾病家族的一部分,包括亨廷顿舞蹈病(HD)。每种病理性蛋白都广泛表达,但CNS内神经元变性的原因仍不清楚。现在许多报告将异常的多聚谷氨酰胺蛋白聚集与发病机制联系起来。先前的一项研究报告称,野生型糖皮质激素受体(wtGR)的激活抑制了源自AR和亨廷顿蛋白的扩增多聚谷氨酰胺蛋白的聚集,而含有内部缺失的突变受体GRDelta108 - 317则增加了多聚谷氨酰胺蛋白的聚集,在这种情况下主要是在细胞核内。在本研究中,我们使用这两种形式的GR在不同的细胞环境中研究扩增的多聚谷氨酰胺AR蛋白。通过细胞生物学和生化方法,我们发现wtGR促进该蛋白的可溶性形式,并防止其在NIH3T3细胞和培养的神经元中发生核聚集。相反,GRDelta108 - 317降低了多聚谷氨酰胺蛋白的溶解度,并在非神经元细胞中导致核聚集体的形成。核聚集体比细胞质聚集体更快地募集hsp72,并且与细胞活力降低有关。有限蛋白酶解和化学交联表明,扩增的AR蛋白独特的可溶性形式是这些不同生物学活性的基础。这些观察结果提供了一个实验框架,以理解为什么扩增的多聚谷氨酰胺蛋白可能仅对某些细胞群体有毒,并表明扩增的多聚谷氨酰胺蛋白独特的蛋白结合或构象可能决定随后的细胞效应,如核定位和细胞毒性。