Maynard Christa J, Böttcher Claudia, Ortega Zaira, Smith Ruben, Florea Bogdan I, Díaz-Hernández Miguel, Brundin Patrik, Overkleeft Hermen S, Li Jia-Yi, Lucas Jose J, Dantuma Nico P
Department of Cell and Molecular Biology, Karolinska Institutet, S-17177 Stockholm, Sweden.
Proc Natl Acad Sci U S A. 2009 Aug 18;106(33):13986-91. doi: 10.1073/pnas.0906463106. Epub 2009 Jul 30.
Aggregation-prone proteins have been suggested to overwhelm and impair the ubiquitin/proteasome system (UPS) in polyglutamine (polyQ) disorders, such as Huntington's disease (HD). Overexpression of an N-terminal fragment of mutant huntingtin (N-mutHtt), an aggregation-prone polyQ protein responsible for HD, obstructs the UPS in cellular models. Furthermore, based on the accumulation of polyubiquitin conjugates in brains of R6/2 mice, which express human N-mutHtt and are one of the most severe polyQ disorder models, it has been proposed that UPS dysfunction is a consistent feature of this pathology, occurring in both in vitro and in vivo models. Here, we have exploited transgenic mice that ubiquitously express a ubiquitin fusion degradation proteasome substrate to directly assess the functionality of the UPS in R6/2 mice or the slower onset R6/1 mice. Although expression of N-mutHtt caused a general inhibition of the UPS in PC12 cells, we did not observe an increase in the levels of proteasome reporter substrate in the brains of R6/2 and R6/1 mice. We show that the increase in ubiquitin conjugates in R6/2 mice can be primarily attributed to an accumulation of large ubiquitin conjugates that are different from the conjugates observed upon UPS inhibition. Together our data show that polyubiquitylated proteins accumulate in R6/2 brain despite a largely operative UPS, and suggest that neurons are able to avoid or compensate for the inhibitory effects of N-mutHtt.
有人提出,在多聚谷氨酰胺(polyQ)疾病(如亨廷顿舞蹈病(HD))中,易于聚集的蛋白质会使泛素/蛋白酶体系统(UPS)不堪重负并造成损害。突变型亨廷顿蛋白(N-mutHtt)的N端片段是一种导致HD的易于聚集的多聚谷氨酰胺蛋白,其过表达会在细胞模型中阻碍UPS。此外,基于R6/2小鼠(表达人类N-mutHtt且是最严重的多聚谷氨酰胺疾病模型之一)大脑中多聚泛素缀合物的积累,有人提出UPS功能障碍是这种病理的一个一致特征,在体外和体内模型中都会发生。在此,我们利用了能普遍表达泛素融合降解蛋白酶体底物的转基因小鼠,以直接评估R6/2小鼠或发病较慢的R6/1小鼠中UPS的功能。虽然N-mutHtt的表达在PC12细胞中导致了UPS的普遍抑制,但我们在R6/2和R6/1小鼠的大脑中未观察到蛋白酶体报告底物水平的增加。我们表明,R6/2小鼠中泛素缀合物的增加主要可归因于与UPS抑制时观察到的缀合物不同的大泛素缀合物的积累。我们的数据共同表明,尽管UPS在很大程度上仍能正常运作,但多聚泛素化蛋白仍在R6/2小鼠大脑中积累,这表明神经元能够避免或补偿N-mutHtt的抑制作用。