泛素连接酶成员FBXO11靶向识别新底物GTF2A1
首发时间:2024-04-24
摘要:目的:研究转录起始因子GTF2A1能否被泛素连接酶成员FBXO11识别并介导其泛素化降解。方法:通过对已发表的转录组和蛋白质组数据整合分析并利用real-time PCR和Western blot验证GTF2A1在FBXO11缺失的红系组细胞系HUDEP-2中的mRNA和蛋白水平。通过免疫共沉淀、蛋白质降解和泛素化水平检测等方法来探究FBXO11对GTF2A1的蛋白稳态和泛素化水平的调控。结果:GTF2A1的蛋白水平在FBXO11缺失的HUDEP-2细胞中出现累积增高,而mRNA水平没有显著改变。FBXO11与GTF2A1存在蛋白质相互作用且FBXO11过表达显著升高GTF2A1泛素化修饰水平并介导其通过泛素蛋白酶体途径发生降解。结论:FBXO11靶向识别新底物GTF2A1,并介导其通过泛素化蛋白酶体途径发生降解。
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E3 Ubiquitin ligase adaptor FBXO11 targets the recognition of the novel substrate GTF2A1
Abstract:Objective: To investigate whether transcription initiation factor GTF2A1 can be recognized by ubiquitin ligase FBXO11. Methods: The mRNA and protein levels of GTF2A1 in FBXO11-deficient HUDEP-2 cells were analyzedby integrating the previous published transcriptome and proteome dataset and verified by using real-time PCR and Western blot. The regulation of ubiquitin ligase FBXO11 on GTF2A1 protein and ubiquitin levels was investigated by immuno-coprecipitation , potein stablility and ubiquitination assays. Results: The protein level of GTF2A1 increased cumulatively in FBXO11-deficient HUDEP-2 cells, but the mRNA level did not change significantly. FBXO11 interacts with GTF2A1 and mediates the degradation of GTF2A1 through ubiquitin proteasome pathway. Conclusion: FBXO11 recognizes and targets the new substrate GTF2A1 for its further degradation via the ubiquitination proteasome pathway.
Keywords: E3 ubiquitin ligase FBXO11 GTF2A1
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