目的筛选影响胆固醇调节元件结合蛋白1a(cholesterol regulatory element binding protein,SREBP1a)蛋白稳定性的去泛素化酶,并探索其调控机制。方法通过去泛素化酶库筛选显著影响SREBP1a表达的去泛素化酶,免疫蛋白印记实验和实时荧光定...目的筛选影响胆固醇调节元件结合蛋白1a(cholesterol regulatory element binding protein,SREBP1a)蛋白稳定性的去泛素化酶,并探索其调控机制。方法通过去泛素化酶库筛选显著影响SREBP1a表达的去泛素化酶,免疫蛋白印记实验和实时荧光定量PCR(qRT-PCR)评估去泛素化酶对SREBP1a以及升脂基因表达的影响;通过红色荧光蛋白标记人源低密度脂蛋白(human Dil-low density lipoprotein,Human Dil-LDL)摄取和油红O染色等实验技术检测细胞摄取低密度脂蛋白(LDL)和脂质沉积情况。结果去泛素化酶库筛选发现泛素特异肽酶37(ubiquitin specific peptidase 37,USP37)可显著增加肝细胞SREBP1a蛋白表达水平,促进胆固醇摄取及脂质沉积。USP37基因敲除可显著降低SREBP1a蛋白表达水平,抑制升脂基因表达及脂质沉积。结论去泛素化酶USP37通过稳定SREBP1a蛋白表达,促进胆固醇摄取及脂质沉积,揭示了SREBP1a翻译后调控的新模式。展开更多
Colorectal cancer(CRC)poses a severe global health challenge with high incidence and mortality rates.USP37 has been identified as the bona fide deubiquitinase of SND1,playing a critical role in stabilizing SND1,thereb...Colorectal cancer(CRC)poses a severe global health challenge with high incidence and mortality rates.USP37 has been identified as the bona fide deubiquitinase of SND1,playing a critical role in stabilizing SND1,thereby augmenting its oncogenic potential.The interaction between USP37 and SND1 was confirmed through extensive proteomics,ubiquitinomics,and interactomics,underscoring their synergistic effects on CRC proliferation and metastasis.Additionally,CDK1 has emerged as a pivotal regulator of USP37,phosphorylating it at threonine 631 rather than serine 628,enhancing its deubiquitinase activity,and consequently stabilizing SND1 to drive CRC malignancy further.Histological analyses of human CRC samples linked the upregulation of CDK1 and USP37 with increased SND1 levels and poor patient prognosis.High-throughput virtual screening and subsequent experimental validation identified Dacarbazine as a pharmacological inhibitor of USP37,and its inhibition disrupted SND1 stability,hindering CRC cell proliferation and metastasis.This study reveals a novel and promising molecular mechanism driving CRC progression through the CDK1-SP37-ND1 axis,highlighting the clinical importance of targeting this pathway to improve patient outcomes.展开更多
文摘目的筛选影响胆固醇调节元件结合蛋白1a(cholesterol regulatory element binding protein,SREBP1a)蛋白稳定性的去泛素化酶,并探索其调控机制。方法通过去泛素化酶库筛选显著影响SREBP1a表达的去泛素化酶,免疫蛋白印记实验和实时荧光定量PCR(qRT-PCR)评估去泛素化酶对SREBP1a以及升脂基因表达的影响;通过红色荧光蛋白标记人源低密度脂蛋白(human Dil-low density lipoprotein,Human Dil-LDL)摄取和油红O染色等实验技术检测细胞摄取低密度脂蛋白(LDL)和脂质沉积情况。结果去泛素化酶库筛选发现泛素特异肽酶37(ubiquitin specific peptidase 37,USP37)可显著增加肝细胞SREBP1a蛋白表达水平,促进胆固醇摄取及脂质沉积。USP37基因敲除可显著降低SREBP1a蛋白表达水平,抑制升脂基因表达及脂质沉积。结论去泛素化酶USP37通过稳定SREBP1a蛋白表达,促进胆固醇摄取及脂质沉积,揭示了SREBP1a翻译后调控的新模式。
基金supported by the National Natural Science Foundation of China(Nos.82103543,82303235,U19A2008 and 82373232)Fundamental Research Funds for the Central Universities(No.WK9110000159,China)+1 种基金the Natural Science Foundation of Anhui Province(No.2108085QH340,China)the China Postdoctoral Science Foundation(No.2021M693082).
文摘Colorectal cancer(CRC)poses a severe global health challenge with high incidence and mortality rates.USP37 has been identified as the bona fide deubiquitinase of SND1,playing a critical role in stabilizing SND1,thereby augmenting its oncogenic potential.The interaction between USP37 and SND1 was confirmed through extensive proteomics,ubiquitinomics,and interactomics,underscoring their synergistic effects on CRC proliferation and metastasis.Additionally,CDK1 has emerged as a pivotal regulator of USP37,phosphorylating it at threonine 631 rather than serine 628,enhancing its deubiquitinase activity,and consequently stabilizing SND1 to drive CRC malignancy further.Histological analyses of human CRC samples linked the upregulation of CDK1 and USP37 with increased SND1 levels and poor patient prognosis.High-throughput virtual screening and subsequent experimental validation identified Dacarbazine as a pharmacological inhibitor of USP37,and its inhibition disrupted SND1 stability,hindering CRC cell proliferation and metastasis.This study reveals a novel and promising molecular mechanism driving CRC progression through the CDK1-SP37-ND1 axis,highlighting the clinical importance of targeting this pathway to improve patient outcomes.
