Ube2V2 Is a Rosetta Stone Bridging Redox and Ubiquitin Codes, Coordinating DNA Damage Responses.
Ube2V2 Is a Rosetta Stone Bridging Redox and Ubiquitin Codes, Coordinating DNA Damage Responses.
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DOI：
10.1021/acscentsci.7b00556
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发表时间：
2018-02-28
影响因子：
18.2
通讯作者：
Aye Y
中科院分区：
文献类型：
--
作者：
Zhao Y;Long MJC;Wang Y;Zhang S;Aye Y
翻译后修饰（ptm）是细胞通信的通用语言。大多数ptm是由酶调控的。然而，亲电药物的重新出现带来了非常规/非酶催化的亲电信号通路的挖掘。尽管在亲电药物设计中利用具有动力学和功能优势的半胱氨酸是最新的推动力，但识别这些传感器仍然具有挑战性。在这里，我们设计了“G-REX”——一种允许活性亲电试剂在体内受控释放的技术。G-REX标记了在真正的kcat/Km条件下结合亲电试剂的第一反应先天半胱氨酸，减轻了与不受控制的大剂量暴露相关的毒性/脱靶效应。G-REX鉴定出两种变链泛素偶联蛋白ube2v1 / ube2v2，它们共享一种新的特权传感器-半胱氨酸。这种非酶催化的ptm触发了对每种蛋白质的特异性反应。因此，G-REX是一种鉴定新型功能性半胱氨酸的无偏方法。与传统的调节靶标活性的活性位点/非活性位点半胱氨酸修饰相比，Ube2V2的修饰变构性地过度激活了其酶活性结合伙伴Ube2N，促进了k63连接的客户端泛素化并刺激了h2axs依赖性DNA损伤反应。这项工作建立了Ube2V2作为连接氧化还原和泛素编码以保护基因组完整性的罗塞塔石碑。G-REX和T-REX双接策略确定了Ube2V2的亲电反应性，从而促进了基因组保护。
Posttranslational modifications (PTMs) are the lingua franca of cellular communication. Most PTMs are enzyme-orchestrated. However, the reemergence of electrophilic drugs has ushered mining of unconventional/non-enzyme-catalyzed electrophile-signaling pathways. Despite the latest impetus toward harnessing kinetically and functionally privileged cysteines for electrophilic drug design, identifying these sensors remains challenging. Herein, we designed “G-REX”—a technique that allows controlled release of reactive electrophiles in vivo. Mitigating toxicity/off-target effects associated with uncontrolled bolus exposure, G-REX tagged first-responding innate cysteines that bind electrophiles under true kcat/Km conditions. G-REX identified two allosteric ubiquitin-conjugating proteins—Ube2V1/Ube2V2—sharing a novel privileged-sensor-cysteine. This non-enzyme-catalyzed-PTM triggered responses specific to each protein. Thus, G-REX is an unbiased method to identify novel functional cysteines. Contrasting conventional active-site/off-active-site cysteine-modifications that regulate target activity, modification of Ube2V2 allosterically hyperactivated its enzymatically active binding-partner Ube2N, promoting K63-linked client ubiquitination and stimulating H2AX-dependent DNA damage response. This work establishes Ube2V2 as a Rosetta-stone bridging redox and ubiquitin codes to guard genome integrity. G-REX and T-REX double tap strategy identifies privileged electrophile responsivity of Ube2V2 to promote genome protection.
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影响因子：
16.6
作者：
通讯作者：
--
影响因子：
15
作者：
Lin HY;Haegele JA;Disare MT;Lin Q;Aye Y
通讯作者：
Aye Y
影响因子：
7.3
作者：
Blewett MM;Xie J;Zaro BW;Backus KM;Altman A;Teijaro JR;Cravatt BF
通讯作者：
Cravatt BF
影响因子：
64.5
作者：
Doil, Carsten;Mailand, Niels;Lukas, Claudia
通讯作者：
Lukas, Claudia
影响因子：
64.5
作者：
Deng, L;Wang, C;Chen, ZJ
通讯作者：
Chen, ZJ