Tait D, Riccio M, Sittler A, Scherzinger E, Santi S, Ognibene A, Maraldi N M, Lehrach H, Wanker E E
Max-Planck Institut für Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin (Dahlem), Germany.
Hum Mol Genet. 1998 Jun;7(6):991-7. doi: 10.1093/hmg/7.6.991.
It has been reported that the ataxin-3 protein containing a polyglutamine sequence in the pathological range (61-84Q) is localized within the nucleus of neuronal cells, whereas ataxin-3 with a normal repeat length (12-37Q) is predominantly a cytoplasmic protein. In this study, the subcellular localization of the full-length ataxin-3 protein with a glutamine sequence in the normal range (Q3KQ22) was analysed in two mammalian cell lines. Using two affinity-purified polyclonal antibodies raised against the N- or C-terminal portion of ataxin-3, the protein was detected predominantly, but not exclusively, in the nucleus of COS-7 as well as neuroblastoma cells by immunofluorescence and confocal laser scanning microscopy (CLSM). The distribution of the protein in these cellular compartments was confirmed by biochemical subcellular fractionations. Furthermore, CLSM revealed that the ataxin-3 protein present in the nucleus of neuroblastoma cells is associated with the inner nuclear matrix. Our results taken together with the finding of a nuclear localization signal in ataxin-3 indicate that the ataxin-3 protein per se translocates to the nucleus and that an expanded glutamine repeat is not essential for this transport.
据报道,含有病理性范围(61 - 84个谷氨酰胺)多谷氨酰胺序列的ataxin-3蛋白定位于神经元细胞核内,而具有正常重复长度(12 - 37个谷氨酰胺)的ataxin-3主要是一种细胞质蛋白。在本研究中,在两种哺乳动物细胞系中分析了具有正常范围谷氨酰胺序列(Q3KQ22)的全长ataxin-3蛋白的亚细胞定位。使用针对ataxin-3的N端或C端部分产生的两种亲和纯化多克隆抗体,通过免疫荧光和共聚焦激光扫描显微镜(CLSM)在COS-7细胞以及神经母细胞瘤细胞的细胞核中主要但并非唯一地检测到该蛋白。通过生化亚细胞分级分离证实了该蛋白在这些细胞区室中的分布。此外,CLSM显示神经母细胞瘤细胞核中存在的ataxin-3蛋白与核内基质相关。我们的结果与在ataxin-3中发现核定位信号一起表明ataxin-3蛋白本身易位至细胞核,并且谷氨酰胺重复序列的扩增对于这种转运并非必不可少。