The Catalytically Inactive Mutation of the Ubiquitin-Conjugating Enzyme CDC34 Affects its Stability and Cell Proliferation
The Catalytically Inactive Mutation of the Ubiquitin-Conjugating Enzyme CDC34 Affects its Stability and Cell Proliferation
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泛素结合酶 CDC34 的催化失活突变影响其稳定性和细胞增殖
DOI：
10.1007/s10930-018-9766-x
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发表时间：
2018-03
期刊：
影响因子：
3
通讯作者：
许国强
中科院分区：
文献类型：
--
作者：
刘勋;张阳;胡展红;李倩;杨陆;许国强
泛素蛋白酶体系统（UPS）在蛋白质稳定性、定位和活性的调节中起着重要作用。由于泛素连接酶E3和去泛素化酶DUB在下游底物识别中的特异性，大量的研究集中在它们的功能上。然而，大多数泛素结合酶E2的作用还没有完全了解，除了它们运输活化的泛素和形成E2-E3蛋白复合物。泛素结合酶CDC 34可以通过UPS促进下游靶标的降解，而其非催化功能仍然是难以捉摸的。在这里，我们发现，催化活性的半胱氨酸丝氨酸（C93 S）的突变导致减少泛素化，增加稳定性，并减弱降解率的CDC 34。通过半定量蛋白质组学，我们确定了CDC 34相互作用的蛋白质，发现野生型和突变型蛋白质有许多差异相互作用的蛋白质。详细研究发现，其中一些参与基因表达、细胞生长和细胞增殖的调节。细胞增殖试验表明，野生型和C93 S蛋白都影响癌细胞系的增殖。数据库分析显示，CDC 34 mRNA在多种癌症中高度表达，这与患者生存率降低相关。这项工作可能有助于阐明这种蛋白质的酶促和非酶促功能，并可能为靶向E2的药物发现提供额外的见解。
The ubiquitin proteasome system (UPS) plays important roles in the regulation of protein stability, localization, and activity. A myriad of studies have focused on the functions of ubiquitin ligases E3s and deubiquitinating enzymes DUBs due to their specificity in the recognition of downstream substrates. However, the roles of the most ubiquitin-conjugating enzymes E2s are not completely understood except that they transport the activated ubiquitin and form E2–E3 protein complexes. Ubiquitin-conjugating enzyme CDC34 can promote the degradation of downstream targets through the UPS whereas its non-catalytic functions are still elusive. Here, we find that mutation of the catalytically active cysteine to serine (C93S) results in the reduced ubiquitination, increased stability, and attenuated degradation rate of CDC34. Through semi-quantitative proteomics, we identify the CDC34-interacting proteins and discover that the wild-type and mutant proteins have many differentially interacted proteins. Detailed examination finds that some of them are involved in the regulation of gene expression, cell growth, and cell proliferation. Cell proliferation assay reveals that both the wild-type and C93S proteins affect the proliferation of a cancer cell line. Database analyses show that CDC34 mRNA is highly expressed in multiple cancers, which is correlated with the reduced patient survival rate. This work may help to elucidate the enzymatic and non-enzymatic functions of this protein and might provide additional insights for drug discovery targeting E2s.
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Cdc34酸性环的抑制剂：一项计算研究，整合了分子动力学，虚拟筛选和对接方法。
影响因子：
2.6
作者：
Arrigoni, Alberto;Bertini, Luca;De Gioia, Luca;Papaleo, Elena
通讯作者：
Papaleo, Elena
DOI：
10.1016/s0021-9258(18)34958-5
发表时间：
1982-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
A. Haas;J. V. Warms;A. Hershko;I. A. Rose
通讯作者：
A. Haas;J. V. Warms;A. Hershko;I. A. Rose
影响因子：
2.9
作者：
F. Gharahdaghi;Catherine R. Weinberg;D. A. Meagher;B. Imai;S. Mische
通讯作者：
F. Gharahdaghi;Catherine R. Weinberg;D. A. Meagher;B. Imai;S. Mische
DOI：
10.1016/s0021-9258(18)34024-9
发表时间：
1982-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
A. Haas;I. A. Rose
通讯作者：
A. Haas;I. A. Rose
蛋白酶体抑制可挽救大脑中与智力低下相关的无义突变
影响因子：
4.8
作者：
Xu, Guoqiang;Jiang, Xiaogang;Jaffrey, Samie R.
通讯作者：
Jaffrey, Samie R.