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K6 linked polyubiquitylation of FADD by CHIP prevents death inducing signaling complex formation suppressing cell death.
Oncogene ( IF 6.9 ) Pub Date : 2018-Sep-01 , DOI: 10.1038/s41388-018-0323-z
Jinho Seo , Eun-Woo Lee , Jihye Shin , Daehyeon Seong , Young Woo Nam , Manhyung Jeong , Seon-Hyeong Lee , Cheolju Lee , Jaewhan Song

Fas-associated death domain (FADD) is an adaptor protein recruiting complexes of caspase 8 to death ligand receptors to induce extrinsic apoptotic cell death in response to a TNF superfamily member. Although, formation of the complex of FADD and caspase 8 upon death stimuli has been studied in detail, posttranslational modifications fine-tuning these processes have yet to be identified. Here we revealed that K6-linked polyubiquitylation of FADD on lysines 149 and 153 mediated by C terminus HSC70-interacting protein (CHIP) plays an important role in preventing formation of the death inducing signaling complex (DISC), thus leading to the suppression of cell death. Cells depleted of CHIP showed higher sensitivity toward death ligands such as FasL and TRAIL, leading to upregulation of DISC formation composed of a death receptor, FADD, and caspase 8. CHIP was able to bind to FADD, induce K6-linked polyubiquitylation of FADD, and suppress DISC formation. By mass spectrometry, lysines 149 and 153 of FADD were found to be responsible for CHIP-mediated FADD ubiquitylation. FADD mutated at these sites was capable of more potent cell death induction as compared with the wild type and was no longer suppressed by CHIP. On the other hand, CHIP deficient in E3 ligase activity was not capable of suppressing FADD function and of FADD ubiquitylation. CHIP depletion in ME-180 cells induced significant sensitization of these cells toward TRAIL in xenograft analyses. These results imply that K6-linked ubiquitylation of FADD by CHIP is a crucial checkpoint in cytokine-dependent extrinsic apoptosis.

中文翻译:

通过CHIP与FADD的K6连接的多泛素化可防止死亡,诱导死亡的信号复合物形成抑制细胞死亡。

Fas相关死亡域(FADD)是一种衔接蛋白,将caspase 8的复合物募集到死亡配体受体上,以响应TNF超家族成员而诱导外在性凋亡细胞死亡。尽管已经详细研究了死亡刺激后FADD和胱天蛋白酶8的复合物的形成,但是尚未确定微调这些过程的翻译后修饰。在这里我们揭示了由C末端HSC70相互作用蛋白(CHIP)介导的赖氨酸149和153上KDD的FADD的K6连接多泛素化在防止死亡诱导信号复合物(DISC)的形成中起重要作用,从而导致细胞抑制死亡。耗尽CHIP的细胞对死亡配体(例如FasL和TRAIL)显示出更高的敏感性,从而导致由死亡受体,FADD和caspase 8组成的DISC形成上调。CHIP能够与FADD结合,诱导K6连接的FADD多聚泛素化,并抑制DISC的形成。通过质谱,发现FADD的赖氨酸149和153负责CHIP介导的FADD的泛素化。与野生型相比,在这些位点突变的FADD能够更有效地诱导细胞死亡,并且不再被CHIP抑制。另一方面,缺乏E3连接酶活性的CHIP不能抑制FADD功能和FADD泛素化。在异种移植分析中,ME-180细胞中的CHIP耗尽诱导了这些细胞对TRAIL的显着敏化。这些结果表明,CHIP的KDD连接的FADD泛素化是细胞因子依赖性外源性细胞凋亡的关键检查点。通过质谱,发现FADD的赖氨酸149和153负责CHIP介导的FADD的泛素化。与野生型相比,在这些位点突变的FADD能够更有效地诱导细胞死亡,并且不再被CHIP抑制。另一方面,缺乏E3连接酶活性的CHIP不能抑制FADD功能和FADD泛素化。在异种移植分析中,ME-180细胞中的CHIP耗尽诱导了这些细胞对TRAIL的显着敏化。这些结果表明,CHIP的KDD连接的FADD泛素化是细胞因子依赖性外源性细胞凋亡的关键检查点。通过质谱,发现FADD的赖氨酸149和153负责CHIP介导的FADD的泛素化。与野生型相比,在这些位点突变的FADD能够更有效地诱导细胞死亡,并且不再被CHIP抑制。另一方面,缺乏E3连接酶活性的CHIP不能抑制FADD功能和FADD泛素化。在异种移植分析中,ME-180细胞中的CHIP耗尽诱导了这些细胞对TRAIL的显着敏化。这些结果表明,CHIP的KDD连接的FADD泛素化是细胞因子依赖性外源性细胞凋亡的关键检查点。与野生型相比,在这些位点突变的FADD能够更有效地诱导细胞死亡,并且不再被CHIP抑制。另一方面,缺乏E3连接酶活性的CHIP不能抑制FADD功能和FADD泛素化。在异种移植分析中,ME-180细胞中的CHIP耗尽诱导了这些细胞对TRAIL的显着敏化。这些结果表明,CHIP的KDD连接的FADD泛素化是细胞因子依赖性外源性细胞凋亡的关键检查点。与野生型相比,在这些位点突变的FADD能够更有效地诱导细胞死亡,并且不再被CHIP抑制。另一方面,缺乏E3连接酶活性的CHIP不能抑制FADD功能和FADD泛素化。在异种移植分析中,ME-180细胞中的CHIP耗尽诱导了这些细胞对TRAIL的显着敏化。这些结果表明,CHIP的KDD连接的FADD泛素化是细胞因子依赖性外源性细胞凋亡的关键检查点。
更新日期:2018-05-23
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