Sayer Jonathan A, Manczak Maria, Akileswaran Lakshmi, Reddy P Hemachandra, Coghlan Vincent M
Neurological Sciences Institute, Oregon Health & Science University, Beaverton, OR 97006, USA.
Neuromolecular Med. 2005;7(4):297-310. doi: 10.1385/NMM:7:4:297.
Although expansion of a polyglutamine tract in the huntingtin protein is known to cause Huntington's disease (HD), there is considerable debate as to how this mutation leads to the selective neuronal loss that characterizes the disease. The observation that mutant huntingtin accumulates in neuronal nuclei has led to the hypothesis that the molecular mechanism may involve the disruption of specific nuclear activities. Recently, several nuclear interaction partners for huntingtin have been identified, including HypA, a splicing factor-like protein of unknown function. Using a yeast two-hybrid screen, we have identified the interaction of HypA with the nuclear scaffold protein NAKAP. Interaction of NAKAP with HypA is specific and occurs both in yeast and in vitro. Deletion-mapping studies indicate that binding occurs via a proline-rich domain in NAKAP with a WW domain of HypA. In cultured cells, NAKAP and HypA localize within the nucleus and copurify with the nuclear matrix. Furthermore, NAKAP associates with HypA from human brain and copurifies with huntingtin protein in brain tissue obtained from HD patients. In HD neurons, NAKAP and mutant huntingtin were colocalized to the nuclear matrix and were found to be components of nuclear aggregates. Hence, the NAKAP-HypA scaffold is a potential nuclear docking site for huntingtin protein and may contribute to the nuclear accumulation of huntingtin observed in HD.
虽然已知亨廷顿蛋白中多聚谷氨酰胺序列的扩展会导致亨廷顿舞蹈症(HD),但对于这种突变如何导致该疾病特有的选择性神经元丧失,仍存在相当大的争议。突变型亨廷顿蛋白在神经元细胞核中积累这一观察结果引发了一种假说,即分子机制可能涉及特定核活动的破坏。最近,已鉴定出几种与亨廷顿蛋白相互作用的核蛋白,包括HypA,一种功能未知的剪接因子样蛋白。通过酵母双杂交筛选,我们鉴定出HypA与核支架蛋白NAKAP之间的相互作用。NAKAP与HypA的相互作用具有特异性,且在酵母和体外均会发生。缺失定位研究表明,结合是通过NAKAP中富含脯氨酸的结构域与HypA的WW结构域发生的。在培养细胞中,NAKAP和HypA定位于细胞核内,并与核基质共同纯化。此外,NAKAP与人脑中的HypA结合,并与从HD患者获得的脑组织中的亨廷顿蛋白共同纯化。在HD神经元中,NAKAP和突变型亨廷顿蛋白共定位于核基质,并被发现是核聚集体的组成部分。因此,NAKAP-HypA支架是亨廷顿蛋白潜在的核停靠位点,可能导致在HD中观察到的亨廷顿蛋白的核积累。