Genome-wide RNA interference screen identifies previously undescribed regulators of polyglutamine aggregation
Genome-wide RNA interference screen identifies previously undescribed regulators of polyglutamine aggregation
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DOI：
10.1073/pnas.0307697101
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发表时间：
2004-04-27
影响因子：
11.1
通讯作者：
Plasterk, RHA
中科院分区：
文献类型：
--
作者：
Nollen, EAA;Garcia, SM;Plasterk, RHA
蛋白质错误折叠和聚集体的形成越来越被认为是人类遗传疾病病理的组成部分，也是许多神经退行性疾病的标志。如多聚谷氨酰胺疾病的例子所示，蛋白质错误折叠的倾向与多聚谷氨酰胺扩张的长度和蛋白质折叠内稳态的年龄依赖性变化有关，这表明蛋白质内稳态缓冲剂的关键作用。为了确定保护细胞免受蛋白质聚集形成的蛋白质因子的补充，我们测试了转基因秀丽隐杆线虫菌株，该菌株在与年龄依赖性蛋白质聚集外观相关的阈值长度处表达聚谷氨酰胺扩增黄色荧光蛋白融合蛋白。我们使用全基因组RNA干扰来识别基因，当被抑制时，会导致蛋白质聚集体的过早出现。我们筛选了186个基因，对应于5类主要的聚谷氨酰胺调节因子：涉及RNA代谢、蛋白质合成、蛋白质折叠和蛋白质降解的基因；以及那些参与蛋白质运输的人。我们认为，每一类都代表一个分子机器，共同组成蛋白质稳态缓冲，响应受损蛋白质的表达，以防止它们的错误折叠和聚集。
Protein misfolding and the formation of aggregates are increasingly recognized components of the pathology of human genetic disease and hallmarks of many neurodegenerative disorders. As exemplified by polyglutamine diseases, the propensity for protein misfolding is associated with the length of polyglutamine expansions and age-dependent changes in protein-folding homeostasis, suggesting a critical role for a protein homeostatic buffer. To identify the complement of protein factors that protects cells against the formation of protein aggregates, we tested transgenic Caenorhabditis elegans strains expressing polyglutamine expansion yellow fluorescent protein fusion proteins at the threshold length associated with the age-dependent appearance of protein aggregation. We used genome-wide RNA interference to identify genes that, when suppressed, resulted in the premature appearance of protein aggregates. Our screen identified 186 genes corresponding to five principal classes of polyglutamine regulators: genes involved in RNA metabolism, protein synthesis, protein folding, and protein degradation; and those involved in protein trafficking. We propose that each of these classes represents a molecular machine collectively comprising the protein homeostatic buffer that responds to the expression of damaged proteins to prevent their misfolding and aggregation.