Boutell J M, Thomas P, Neal J W, Weston V J, Duce J, Harper P S, Jones A L
Institute of Medical Genetics, University of Wales College of Medicine, Cardiff, UK.
Hum Mol Genet. 1999 Sep;8(9):1647-55. doi: 10.1093/hmg/8.9.1647.
We detected an interaction of the N-terminus of huntingtin (htt171) with the C-terminal region of the nuclear receptor co-repressor (N-CoR) using the yeast two-hybrid system. This interaction was repeat length dependent and specific to htt171; the co-repressor did not interact with the repeat carrying a section of atrophin 1 nor with the androgen receptor or polyglutamine alone. The interaction was confirmed using His-tagged Escherichia coli -expressed C-terminal human and rat co-repressor protein which pulled full-length huntingtin out of homogenized rat brain and in pull-down assays. The N-CoR represses transcription from sequence-specific ligand-activated receptors such as the retinoid X-thyroid hormone receptor dimers and other nuclear receptors including Mad-Max receptor dimers. The mechanism of this repression appears to be through the formation of a complex of repressor proteins including the N-CoR, mSin3 and histone deacetylases. We have used N-CoR and mSin3A antibodies in immunohistochemical studies and find that in Huntington's disease (HD) cortex and caudate, the cellular localization of these proteins is exclusively cytoplasmic whilst in control brain they are localized in the nucleus as well as the cytoplasm; mSin3A immunoreactivity also occurred in a subset of huntingtin positive intranuclear inclusions. The relocalization of repressor proteins in HD brain may alter transcription and be involved in the pathology of the disease.
我们使用酵母双杂交系统检测到亨廷顿蛋白(htt171)的N端与核受体共抑制因子(N-CoR)的C端区域存在相互作用。这种相互作用依赖于重复序列长度,且对htt171具有特异性;共抑制因子既不与携带一段萎缩素1的重复序列相互作用,也不与雄激素受体或单独的聚谷氨酰胺相互作用。使用带His标签的、在大肠杆菌中表达的人源和大鼠源C端共抑制因子蛋白在下拉实验中从匀浆的大鼠脑中拉出全长亨廷顿蛋白,从而证实了这种相互作用。N-CoR抑制来自序列特异性配体激活受体(如视黄酸X-甲状腺激素受体二聚体)以及包括Mad-Max受体二聚体在内的其他核受体的转录。这种抑制机制似乎是通过形成包括N-CoR、mSin3和组蛋白去乙酰化酶在内的抑制蛋白复合物来实现的。我们在免疫组织化学研究中使用了N-CoR和mSin3A抗体,发现在亨廷顿病(HD)的皮质和尾状核中,这些蛋白的细胞定位仅在细胞质中,而在对照脑中它们同时定位于细胞核和细胞质中;mSin3A免疫反应性也出现在一部分亨廷顿蛋白阳性的核内包涵体中。HD脑中抑制蛋白的重新定位可能会改变转录并参与该疾病的病理过程。