Chung Fu-Zon, Sahasrabuddhe Anagh A, Ma Kaiyu, Chen Xiaofei, Basrur Venkatesha, Lim Megan S, Elenitoba-Johnson Kojo S J
From the Department of Pathology.
From the Department of Pathology, the Center for Computational Medicine and Bioinformatics, and.
J Biol Chem. 2014 Oct 10;289(41):28448-59. doi: 10.1074/jbc.M114.561241. Epub 2014 Aug 20.
Fbxo45 is an atypical E3 ubiquitin ligase, which specifically targets proteins for ubiquitin-mediated degradation. Fbxo45 ablation results in defective neuronal differentiation and abnormal formation of neural connections; however, the mechanisms underlying these defects are poorly understood. Using an unbiased mass spectrometry-based proteomic screen, we show here that N-cadherin is a novel interactor of Fbxo45. N-cadherin specifically interacts with Fbxo45 through two consensus motifs overlapping the site of calcium-binding and dimerization of the cadherin molecule. N-cadherin interaction with Fbxo45 is significantly abrogated by calcium treatment. Surprisingly, Fbxo45 depletion by RNAi-mediated silencing results in enhanced proteolysis of N-cadherin. Conversely, ectopic expression of Fbxo45 results in decreased proteolysis of N-cadherin. Fbxo45 depletion results in dramatic reduction in N-cadherin expression, impaired neuronal differentiation, and diminished formation of neuronal processes. Our studies reveal an unanticipated role for an F-box protein that inhibits proteolysis in the regulation of a critical biological process.
Fbxo45是一种非典型的E3泛素连接酶,它特异性地靶向蛋白质进行泛素介导的降解。Fbxo45缺失会导致神经元分化缺陷和神经连接异常形成;然而,这些缺陷背后的机制尚不清楚。通过基于无偏差质谱的蛋白质组学筛选,我们在此表明N-钙黏蛋白是Fbxo45的一种新型相互作用蛋白。N-钙黏蛋白通过与钙黏蛋白分子的钙结合和二聚化位点重叠的两个共有基序与Fbxo45特异性相互作用。钙处理会显著消除N-钙黏蛋白与Fbxo45的相互作用。令人惊讶的是,RNAi介导的沉默导致Fbxo45缺失会增强N-钙黏蛋白的蛋白水解。相反,Fbxo45的异位表达会导致N-钙黏蛋白的蛋白水解减少。Fbxo45缺失会导致N-钙黏蛋白表达显著降低、神经元分化受损以及神经元突起形成减少。我们的研究揭示了一种F-box蛋白在抑制蛋白水解以调节关键生物学过程中的意外作用。