title: Proteomic analysis of trichloroethylene-induced alterations in expression, distribution, and interactions of SET/TAF-Iα and two SET/TAF-Iα-binding proteins, eEF1A1 and eEF1A2, in hepatic L-02 cells.
abstract: Emerging evidence indicates that trichloroethylene (TCE) exposure causes severe hepatotoxicity. However, the mechanisms of TCE hepatotoxicity remain unclear. Recently, we reported that TCE exposure up-regulated the expression of the oncoprotein SET/TAF-Iα and SET knockdown attenuated TCE-induced cytotoxicity in hepatic L-02 cells. To decipher the function of SET/TAF-Iα and its contributions to TCE-induced hepatotoxicity, we employed a proteomic analysis of SET/TAF-Iα with tandem affinity purification to identify SET/TAF-Iα-binding proteins. We identified 42 novel Gene Ontology co-annotated SET/TAF-Iα-binding proteins. The identifications of two of these proteins (eEF1A1, elongation factor 1-alpha 1; eEF1A2, elongation factor 1-alpha 2) were confirmed by Western blot analysis and co-immunoprecipitation (Co-IP). Furthermore, we analyzed the effects of TCE on the expression, distribution and interactions of eEF1A1, eEF1A2 and SET in L-02 cells. Western blot analysis reveals a significant up-regulation of eEF1A1, eEF1A2 and two isoforms of SET, and immunocytochemical analysis reveals that eEF1A1 and SET is redistributed by TCE. SET is redistributed from the nucleus to the cytoplasm, while eFE1A1 is translocated from the cytoplasm to the nucleus. Moreover, we find by Co-IP that TCE exposure significantly increases the interaction of SET with eEF1A2. Our data not only provide insights into the physiological functions of SET/TAF-Iα and complement the SET interaction networks, but also demonstrate that TCE exposure induces alterations in the expression, distribution and interactions of SET and its binding partners. These alterations may constitute the mechanisms of TCE cytotoxicity.
authors: Key Laboratory of Modern Toxicology of Shenzhen, Medical Key Laboratory of GuangDong Province, Medical Key Laboratory of Health Toxicology of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.
authors: Wen-Xu
authors: WX
authors: Hong
authors: Liang
authors: L
authors: Yang
authors: Moutong
authors: M
authors: Chen
authors: Xifei
authors: X
authors: Yang
authors: Xiaohu
authors: X
authors: Ren
authors: Shisong
authors: S
authors: Fang
authors: Jinbo
authors: J
authors: Ye
authors: Haiyan
authors: H
authors: Huang
authors: Chaoqiong
authors: C
authors: Peng
authors: Li
authors: L
authors: Zhou
authors: Xinfeng
authors: X
authors: Huang
authors: Fan
authors: F
authors: Yang
authors: Desheng
authors: D
authors: Wu
authors: Zhixiong
authors: Z
authors: Zhuang
authors: Jianjun
authors: J
authors: Liu
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D004268:DNA-Binding Proteins; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology; D056488:Histone Chaperones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D040901:Proteomics / Q000379:methods; D012997:Solvents / Q000633:toxicity*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014241:Trichloroethylene / Q000633:toxicity*; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: D004268:DNA-Binding Proteins; C094814:EEF1A1 protein, human; C503234:EEF1A2 protein, human; D056488:Histone Chaperones; D020648:Peptide Elongation Factor 1; C513473:SET protein, human; D012997:Solvents; D014157:Transcription Factors; D014241:Trichloroethylene