Nagai Y, Onodera O, Chun J, Strittmatter W J, Burke J R
Department of Medicine (Neurology), Duke University Medical Center, Durham, North Carolina, 27710, USA.
Exp Neurol. 1999 Feb;155(2):195-203. doi: 10.1006/exnr.1998.6991.
Eight inherited neurodegenerative diseases are caused by genes with expanded CAG repeats coding for polyglutamine domains in the disease-producing proteins. The mechanism by which this expanded polyglutamine domain causes neurodegenerative disease is unknown, but nuclear and cytoplasmic polyglutamine protein aggregation is a common feature. In transfected COS7 cells, expanded polyglutamine proteins aggregate and disrupt the vimentin intermediate filament network. Since neurons have an intermediate filament network composed of neurofilament (NF) and NF abnormalities occur in neurodegenerative diseases, we examined whether pathologic-length polyglutamine domain proteins also interact with NF. We expressed varying lengths polyglutamine-green fluorescent protein fusion proteins in a neuroblast cell line, TR1. Pathologic-length polyglutamine-GFP fusion proteins formed large cytoplasmic aggregates surrounded by neurofilament. Immunoisolation of pathologic-length polyglutamine proteins coisolated 68-kDa NF protein demonstrating molecular interaction. These observations suggest that polyglutamine interaction with NF is important in the pathogenesis of the polyglutamine repeat diseases.
八种遗传性神经退行性疾病是由基因中编码致病蛋白中多聚谷氨酰胺结构域的CAG重复序列扩增引起的。这种扩增的多聚谷氨酰胺结构域导致神经退行性疾病的机制尚不清楚,但核内和胞质内多聚谷氨酰胺蛋白聚集是一个共同特征。在转染的COS7细胞中,扩增的多聚谷氨酰胺蛋白聚集并破坏波形蛋白中间丝网络。由于神经元具有由神经丝(NF)组成的中间丝网络,且神经退行性疾病中会出现NF异常,我们研究了病理长度的多聚谷氨酰胺结构域蛋白是否也与NF相互作用。我们在神经母细胞瘤细胞系TR1中表达了不同长度的多聚谷氨酰胺-绿色荧光蛋白融合蛋白。病理长度的多聚谷氨酰胺-GFP融合蛋白形成了被神经丝包围的大的胞质聚集体。对病理长度的多聚谷氨酰胺蛋白进行免疫分离时,共分离出68 kDa的NF蛋白表明存在分子相互作用。这些观察结果表明,多聚谷氨酰胺与NF的相互作用在多聚谷氨酰胺重复疾病的发病机制中很重要。