Potentiated Hsp104 variants suppress toxicity of diverse neurodegenerative disease-linked proteins
Potentiated Hsp104 variants suppress toxicity of diverse neurodegenerative disease-linked proteins
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DOI：
10.1242/dmm.016113
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发表时间：
2014-10-01
影响因子：
4.3
通讯作者：
Shorter, James
中科院分区：
文献类型：
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作者：
Jackrel, Meredith E.;Shorter, James
蛋白质错误折叠与许多致命性神经退行性疾病有关，包括肌萎缩侧索硬化症(ALS)和帕金森病(PD)。没有治疗方法可以逆转这些蛋白质错误折叠事件。我们的目标是应用Hsp104，一种来自酵母的六聚体AAA+蛋白，来靶向错误折叠的构象以重新激活。HSP104可溶解无序的聚集体和淀粉样蛋白，但对人类神经退行性疾病蛋白的活性有限。因此，我们之前已经设计出了增强的Hsp104变体，它可以抑制酿酒酵母中ALS和PD蛋白的聚集、蛋白毒性和恢复正确的蛋白质定位，并减轻动物PD模型中的神经退行性变。在这里，我们建立了增强的HSP104变异体具有广泛的底物特异性，并在酵母中抑制野生型TDP-43、FUS和α-突触核蛋白诱导的毒性和聚集，以及导致神经退行性疾病的这些蛋白的错义突变版本。增强的HSP104变体也可以挽救TAF15的毒性和聚集性，但不能挽救EWSR1，这两种具有Pron样结构域的RNA结合蛋白与ALS和额颞痴呆的发展有关。因此，增强的Hsp104变异体并不完全是非特异性的。事实上，它们并不只是展示任何天然折叠的蛋白质。相反，增强的Hsp104变体被微调以展开含有野生型Hsp104不能识别的短无结构束的蛋白质。我们的研究确立了增强的HSP104变异体的广泛用途。
Protein misfolding is implicated in numerous lethal neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS) and Parkinson disease (PD). There are no therapies that reverse these protein-misfolding events. We aim to apply Hsp104, a hexameric AAA+ protein from yeast, to target misfolded conformers for reactivation. Hsp104 solubilizes disordered aggregates and amyloid, but has limited activity against human neurodegenerative disease proteins. Thus, we have previously engineered potentiated Hsp104 variants that suppress aggregation, proteotoxicity and restore proper protein localization of ALS and PD proteins in Saccharomyces cerevisiae, and mitigate neurodegeneration in an animal PD model. Here, we establish that potentiated Hsp104 variants possess broad substrate specificity and, in yeast, suppress toxicity and aggregation induced by wild-type TDP-43, FUS and alpha-synuclein, as well as missense mutant versions of these proteins that cause neurodegenerative disease. Potentiated Hsp104 variants also rescue toxicity and aggregation of TAF15 but not EWSR1, two RNA-binding proteins with a prion-like domain that are connected with the development of ALS and frontotemporal dementia. Thus, potentiated Hsp104 variants are not entirely non-specific. Indeed, they do not unfold just any natively folded protein. Rather, potentiated Hsp104 variants are finely tuned to unfold proteins bearing short unstructured tracts that are not recognized by wild-type Hsp104. Our studies establish the broad utility of potentiated Hsp104 variants.