Departments of Molecular and Systems Biology, Geisel School of Medicine at Dartmouth College, Hanover, New Hampshire, 03755, United States.
Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth College, Hanover, New Hampshire, 03755, United States.
Sci Rep. 2018 Jun 22;8(1):9554. doi: 10.1038/s41598-018-27900-5.
Huntington's disease is caused by the pathological expansion of a polyglutamine (polyQ) stretch in Huntingtin (Htt), but the molecular mechanisms by which polyQ expansion in Htt causes toxicity in selective neuronal populations remain poorly understood. Interestingly, heterologous expression of expanded polyQ Htt is toxic in Saccharomyces cerevisiae cells, but has no effect in Schizosaccharomyces pombe, a related yeast species possessing very few endogenous polyQ or Q/N-rich proteins. Here, we used a comprehensive and unbiased mass spectrometric approach to identify proteins that bind Htt in a length-dependent manner in both species. Analysis of the expanded polyQ-associated proteins reveals marked enrichment of proteins that are localized to and play functional roles in nucleoli and mitochondria in S. cerevisiae, but not in S. pombe. Moreover, expanded polyQ Htt appears to interact preferentially with endogenous polyQ and Q/N-rich proteins, which are rare in S. pombe, as well as proteins containing coiled-coil motifs in S. cerevisiae. Taken together, these results suggest that polyQ expansion of Htt may cause cellular toxicity in S. cerevisiae by sequestering endogenous polyQ and Q/N-rich proteins, particularly within nucleoli and mitochondria.
亨廷顿病是由亨廷顿蛋白(Htt)中的病理性聚谷氨酰胺(polyQ)延伸引起的,但聚谷氨酰胺延伸在 Htt 中如何导致选择性神经元群体毒性的分子机制仍知之甚少。有趣的是,在酿酒酵母细胞中,扩增的 polyQ Htt 的异源表达是有毒的,但在亲缘酵母物种裂殖酵母中却没有影响,裂殖酵母中很少有内源性 polyQ 或 Q/N 丰富蛋白。在这里,我们使用了一种全面而无偏的质谱方法,在这两种物种中以长度依赖性的方式鉴定与 Htt 结合的蛋白质。对扩增的 polyQ 相关蛋白的分析表明,在酿酒酵母中,定位于核仁并在核仁中发挥功能作用的蛋白以及在核仁中发挥功能作用的蛋白明显富集,但在裂殖酵母中则没有。此外,扩增的 polyQ Htt 似乎更倾向于与内源性 polyQ 和 Q/N 丰富蛋白相互作用,而这些蛋白在裂殖酵母中很少见,而在酿酒酵母中则与含有卷曲螺旋结构域的蛋白相互作用。综上所述,这些结果表明,Htt 的 polyQ 延伸可能通过隔离内源性 polyQ 和 Q/N 丰富蛋白,特别是在核仁和线粒体中,导致酿酒酵母的细胞毒性。