Assessment of SENP3-interacting proteins in hepatocytes treated with diethylnitrosamine by BioID assay  

作　　者：Fei Chen Hongyu Yan Chu Guo Huiqin Zhu Jing Yi Xuxu Sun Jie Yang 

机构地区：[1]Department of Biochemistry and Molecular Cell Biology,Shanghai Key Laboratory for Tumor Microenvironment and Inflammation,Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China

出　　处：《Acta Biochimica et Biophysica Sinica》2021年第9期1237-1246,共10页生物化学与生物物理学报（英文版）

基　　金：the grants from the National Natural Science Foundation of China(No.31771522);Shanghai Municipal Science and Technology Commission(No.16ZR1418400);the Innovative Research Team of High-level Local Universities in Shanghai,the National Natural Science Foundation of China(No.31970587 to Dr Xuxu Sun);the Foundation of Shanghai Oriental Scholar(No.TP2018045).

摘　　要：SUMOylation of proteins regulates cell behaviors and is reversibly removed by small ubiquitin-like modifier(SUMO)-specific proteases(SENPs).The SENP family member SENP3 is involved in SUMO2/3 deconjugation and has been reported to sense cell stress and accumulate in several human cancer cells and macrophages.We previously reported that Senp3-knockout heterozygous mice showed smaller liver,but the pertinent mechanisms of SENP3 and SUMOylated substrates remain unclear.Thus,in this study,we investigated the interacting proteins with SENP3 and the alteration in hepatocytes treated with the xenobiotic diethylnitrosamine(DEN),which is specifically transformed in the liver and induces DNA double-strand breaks.Our data revealed that a certain amount of SENP3 was present in normal,untreated hepatocytes;however,DEN treatment promoted rapid SENP3 accumulation.SENP3 was mainly localized in the nuclei,and its level was significantly increased in the cytoplasm after 2 h of DEN treatment.The application of the recent proximity-dependent biotinylation(BioID)method led to the identification of 310 SENP3-interacting proteins that were involved in not only gene transcription but also RNA splicing,protein folding,and metabolism.Furthermore,after DEN exposure for a short duration,ribosomal proteins as well as proteins associated with mitochondrial ATP synthesis,membrane transport,and bile acid synthesis,rather than DNA repair proteins,were identified.This study provides insights into the diverse regulatory roles of SENP3,and the BioID method seems to be efficient for identifying physiologically relevant insoluble proteins.

关 键 词：SUMO specific peptidase 3 SUMOYLATION BioID HEPATOCYTES DIETHYLNITROSAMINE 

分 类 号：R575[医药卫生—消化系统]