Natural Genetic Variation in Yeast Reveals That NEDD4 Is a Conserved Modifier of Mutant Polyglutamine Aggregation.
Natural Genetic Variation in Yeast Reveals That NEDD4 Is a Conserved Modifier of Mutant Polyglutamine Aggregation.
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酵母中的自然遗传变异表明NEDD4是突变聚谷氨酸聚集的保守修饰剂。
DOI：
10.1534/g3.118.200289
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发表时间：
2018-11-06
期刊：
影响因子：
--
通讯作者：
Hughes RE
中科院分区：
文献类型：
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作者：
Peters TW;Nelson CS;Gerencser AA;Dumas KJ;Tavshanjian B;Chang KC;Lithgow GJ;Hughes RE
晚发性蛋白质病的一个共同特征是有毒蛋白质构象异构体和聚集体在受影响的组织中的积累。在寻找潜在的药物靶点时，许多研究使用高通量筛选来寻找改变错误折叠蛋白质细胞毒性的基因。这种方法的补充是专注于使用蛋白质聚集作为表型读数的策略，以确定控制聚集体形成和维持的途径。在这里，我们使用芽殖酵母菌株之间的自然变异来遗传地绘制基因座，这些基因座影响来自亨廷顿蛋白突变形式的含聚谷氨酰胺蛋白的聚集，亨廷顿蛋白是亨廷顿病的病原体。来自野生和实验室酵母菌株之间的杂交后代的连锁分析揭示了两个多态位点，修改聚谷氨酰胺聚集。一个基因座含有基因RFU 1，其修饰由E3-泛素连接酶复合物Rsp 5靶向的错误折叠蛋白质的泛素化状态。Rsp 5复合物和哺乳动物同源物NEDD 4的活性在响应蛋白质组应激而维持蛋白质稳态中是至关重要的。我们的分析还表明，连锁的聚集表型的一个独特的基因座含有一个基因编码的Rsp 5相互作用的Bul 2蛋白。等位基因交换实验验证了RFU 1和BUL 2对亨廷顿蛋白聚集的影响。此外，我们发现线虫秀丽隐杆线虫的Rsp 5的直系同源物，wwp-1，也负调节聚谷氨酰胺聚集。人细胞中NEDD 4的敲低同样改变了聚谷氨酰胺聚集。两者合计，这些结果涉及保守的过程，涉及泛素调节网络，修改蛋白质聚集，并提供新的治疗目标聚谷氨酰胺和其他蛋白质折叠疾病。
A feature common to late onset proteinopathic disorders is an accumulation of toxic protein conformers and aggregates in affected tissues. In the search for potential drug targets, many studies used high-throughput screens to find genes that modify the cytotoxicity of misfolded proteins. A complement to this approach is to focus on strategies that use protein aggregation as a phenotypic readout to identify pathways that control aggregate formation and maintenance. Here we use natural variation between strains of budding yeast to genetically map loci that influence the aggregation of a polyglutamine-containing protein derived from a mutant form of huntingtin, the causative agent in Huntington disease. Linkage analysis of progeny derived from a cross between wild and laboratory yeast strains revealed two polymorphic loci that modify polyglutamine aggregation. One locus contains the gene RFU1 which modifies ubiquitination states of misfolded proteins targeted by the E3-ubiquitin ligase complex Rsp5. Activity of the Rsp5 complex, and the mammalian homolog NEDD4, are critical in maintaining protein homeostasis in response to proteomic stress. Our analysis also showed linkage of the aggregation phenotype to a distinct locus containing a gene encoding the Rsp5-interacting Bul2 protein. Allele-swap experiments validated the impact of both RFU1 and BUL2 on huntingtin aggregation. Furthermore, we found that the nematode Caenorhabditis elegans’ ortholog of Rsp5, wwp-1, also negatively regulates polyglutamine aggregation. Knockdown of the NEDD4 in human cells likewise altered polyglutamine aggregation. Taken together, these results implicate conserved processes involving the ubiquitin regulation network that modify protein aggregation and provide novel therapeutic targets for polyglutamine and other protein folding diseases.