Center for Human Genetic Research, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA.
Hum Mol Genet. 2010 Feb 15;19(4):573-83. doi: 10.1093/hmg/ddp524. Epub 2009 Nov 23.
Huntington's disease (HD) is caused by expansion of the polymorphic polyglutamine segment in the huntingtin protein. Full-length huntingtin is thought to be a predominant HEAT repeat alpha-solenoid, implying a role as a facilitator of macromolecular complexes. Here we have investigated huntingtin's domain structure and potential intersection with epigenetic silencer polycomb repressive complex 2 (PRC2), suggested by shared embryonic deficiency phenotypes. Analysis of a set of full-length recombinant huntingtins, with different polyglutamine regions, demonstrated dramatic conformational flexibility, with an accessible hinge separating two large alpha-helical domains. Moreover, embryos lacking huntingtin exhibited impaired PRC2 regulation of Hox gene expression, trophoblast giant cell differentiation, paternal X chromosome inactivation and histone H3K27 tri-methylation, while full-length endogenous nuclear huntingtin in wild-type embryoid bodies (EBs) was associated with PRC2 subunits and was detected with trimethylated histone H3K27 at Hoxb9. Supporting a direct stimulatory role, full-length recombinant huntingtin significantly increased the histone H3K27 tri-methylase activity of reconstituted PRC2 in vitro, and structure-function analysis demonstrated that the polyglutamine region augmented full-length huntingtin PRC2 stimulation, both in Hdh(Q111) EBs and in vitro, with reconstituted PRC2. Knowledge of full-length huntingtin's alpha-helical organization and role as a facilitator of the multi-subunit PRC2 complex provides a novel starting point for studying PRC2 regulation, implicates this chromatin repressive complex in a neurodegenerative disorder and sets the stage for further study of huntingtin's molecular function and the impact of its modulatory polyglutamine region.
亨廷顿病(HD)是由亨廷顿蛋白中多态性聚谷氨酰胺片段的扩展引起的。全长亨廷顿被认为是主要的热重复α-螺旋,暗示其作为大分子复合物的促进剂的作用。在这里,我们研究了亨廷顿的结构域结构及其与表观遗传沉默多梳抑制复合物 2(PRC2)的潜在交叉,这是由共同的胚胎缺陷表型所暗示的。对一组具有不同聚谷氨酰胺区的全长重组亨廷顿进行分析,结果表明其具有显著的构象灵活性,具有可接近的铰链将两个大的α-螺旋结构域分开。此外,缺乏亨廷顿的胚胎表现出 PRC2 对 Hox 基因表达、滋养层巨细胞分化、父本 X 染色体失活和组蛋白 H3K27 三甲基化的调节受损,而野生型胚状体(EBs)中的全长内源性核亨廷顿与 PRC2 亚基相关联,并在 Hoxb9 处检测到组蛋白 H3K27 三甲基化。支持直接刺激作用,全长重组亨廷顿显著增加了体外重构的 PRC2 的组蛋白 H3K27 三甲基化酶活性,结构-功能分析表明,聚谷氨酰胺区增强了全长亨廷顿 PRC2 的刺激作用,无论是在 Hdh(Q111)EBs 中还是在体外,与重构的 PRC2 一起。全长亨廷顿的α-螺旋组织和作为多亚基 PRC2 复合物促进剂的作用的知识为研究 PRC2 调节提供了一个新的起点,暗示该染色质抑制复合物参与神经退行性疾病,并为进一步研究亨廷顿的分子功能及其调节性聚谷氨酰胺区的影响奠定了基础。