The Ubiquitin Proteasome System Is a Key Regulator of Pluripotent Stem Cell Survival and Motor Neuron Differentiation

被引:36
作者
Bax, Monique [1 ,2 ]
McKenna, Jessie [3 ]
Do-Ha, Dzung [1 ,2 ]
Stevens, Claire H. [1 ,2 ]
Higginbottom, Sarah [1 ,2 ]
Balez, Rachelle [1 ,2 ]
Cabral-da-Silva, Mauricio E. Castro [1 ,2 ]
Farrawell, Natalie E. [1 ,2 ]
Engel, Martin [1 ,2 ]
Poronnik, Philip [4 ]
Yerbury, Justin J. [1 ,2 ]
Saunders, Darren N. [3 ]
Ooi, Lezanne [1 ,2 ]
机构
[1] Illawarra Hlth & Med Res Inst, Northfields Ave, Wollongong, NSW 2522, Australia
[2] Univ Wollongong, Sch Chem & Mol Biosci, Northfields Ave, Wollongong, NSW 2522, Australia
[3] Univ New South Wales, Sch Med Sci, Sydney, NSW 2052, Australia
[4] Univ Sydney, Fac Med & Hlth, Sch Med Sci, Camperdown, NSW 2050, Australia
基金
英国医学研究理事会; 澳大利亚国家健康与医学研究理事会;
关键词
ubiquitin; ubiquitinomics; UBA1; induced pluripotent stem cell; motor neuron; motor neurone disease; amyotrophic lateral sclerosis; ACTIVATING ENZYME E1; ALS; MUTATIONS; DEGENERATION; GENERATION; INHIBITORS; MODELS; TOOLS; LINES;
D O I
10.3390/cells8060581
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The ubiquitin proteasome system (UPS) plays an important role in regulating numerous cellular processes, and a dysfunctional UPS is thought to contribute to motor neuron disease. Consequently, we sought to map the changing ubiquitome in human iPSCs during their pluripotent stage and following differentiation to motor neurons. Ubiquitinomics analysis identified that spliceosomal and ribosomal proteins were more ubiquitylated in pluripotent stem cells, whilst proteins involved in fatty acid metabolism and the cytoskeleton were specifically ubiquitylated in the motor neurons. The UPS regulator, ubiquitin-like modifier activating enzyme 1 (UBA1), was increased 36-fold in the ubiquitome of motor neurons compared to pluripotent stem cells. Thus, we further investigated the functional consequences of inhibiting the UPS and UBA1 on motor neurons. The proteasome inhibitor MG132, or the UBA1-specific inhibitor PYR41, significantly decreased the viability of motor neurons. Consistent with a role of the UPS in maintaining the cytoskeleton and regulating motor neuron differentiation, UBA1 inhibition also reduced neurite length. Pluripotent stem cells were extremely sensitive to MG132, showing toxicity at nanomolar concentrations. The motor neurons were more resilient to MG132 than pluripotent stem cells but demonstrated higher sensitivity than fibroblasts. Together, this data highlights the important regulatory role of the UPS in pluripotent stem cell survival and motor neuron differentiation.
引用
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页数:17
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