From the Department of Pharmacology, College of Pharmaceutical Sciences, Jiangsu Key Laboratory of Translational Research for Neuro-Psycho-Diseases, Soochow University, Suzhou, Jiangsu 215123, China and.
J Biol Chem. 2013 Oct 11;288(41):29573-85. doi: 10.1074/jbc.M113.472092. Epub 2013 Aug 27.
A nonsense mutation in cereblon (CRBN) causes autosomal recessive nonsyndromic mental retardation. Cereblon is a substrate receptor for the Cullin-RING E3 ligase complex and couples the ubiquitin ligase to specific ubiquitination targets. The CRBN nonsense mutation (R419X) results in a protein lacking 24 amino acids at its C terminus. Although this mutation has been linked to mild mental retardation, the mechanism by which the mutation affects CRBN function is unknown. Here, we used biochemical and mass spectrometric approaches to explore the function of this mutant. We show that the protein retains its ability to assemble into a Cullin-RING E3 ligase complex and catalyzes the ubiquitination of CRBN-target proteins. However, we find that this mutant exhibits markedly increased levels of autoubiquitination and is more readily degraded by the proteasome than the wild type protein. We also show that the level of the mutant protein can be restored by a treatment of cells with a clinically utilized proteasome inhibitor, suggesting that this agent may be useful for the treatment of mental retardation associated with the CRBN R419X mutation. These data demonstrate that enhanced autoubiquitination and degradation account for the defect in CRBN activity that leads to mental retardation.
cereblon(CRBN)中的无意义突变导致常染色体隐性非综合征性智力低下。 cereblon 是 Cullin-RING E3 连接酶复合物的底物受体,将泛素连接酶与特定的泛素化靶标连接起来。CRBN 无意义突变(R419X)导致其 C 末端缺失 24 个氨基酸。尽管该突变与轻度智力低下有关,但突变影响 CRBN 功能的机制尚不清楚。在这里,我们使用生化和质谱方法来探索该突变体的功能。我们表明,该蛋白保留其组装成 Cullin-RING E3 连接酶复合物的能力,并催化 CRBN 靶蛋白的泛素化。然而,我们发现该突变体表现出明显增加的自身泛素化水平,并且比野生型蛋白更容易被蛋白酶体降解。我们还表明,细胞用临床应用的蛋白酶体抑制剂处理可以恢复突变蛋白的水平,表明该药物可能对治疗与 CRBN R419X 突变相关的智力低下有用。这些数据表明,增强的自身泛素化和降解导致了导致智力低下的 CRBN 活性缺陷。