中国病理生理杂志2012，Vol.28Issue(5)：870-877,8.DOI:10.3969/j.issn.1000-4718.2012.05.018
酵母双杂交技术鉴定FAT10新的结合蛋白:eEF1A1
Identification of a novel FAT10-binding protein, eEF1A1, by a yeast two-hybrid system
摘要
Abstract
AIM: To invesligale the role of HLA - F adjacent transcript 10 (FAT10) in tumorigenesis. METHODS; GAL4 yeast Lwo - hybrid assay was performed to screen the human Hep3B hepatoma cell cDNA library for obtaining the host cell prolein molecules which inleracl with FAT10. The immunofluoresence co - localization and co - im-munoprecipitation assay were applied to confirm the inleraclion of the idenlified proleins. FAT10 was knockdown by siRNA lo invesligale the changes of eukaryolic translation elongalion faclor 1 alpha 1 (eEFlAl) expression. RESULTS; The eEFlAl was selected from the hosl cells using yeasl lwo - hybrid assay. The results of co- localizalion and co - immuno-precipilalion assays further confirmed the inleraclion belween FAT10 and eEFlAl. Interestingly, when FAT10 was down -regulaled by siRNA in Hep3B cells, the prolein expression of endogenous eEFlAl was also found lo be significantly repressed. Furthermore, the expression of FAT10 was reduced by siRNA knockdown, thus resulting in the down - regulation of eEFlAl expression at both mRNA and prolein levels in MHCC97H cells. CONCLUSION: We propose a model in which eEFlAl serves as a substrale of FAT10 to accomplish, in part, its function in regulating the biological behaviors of tumor cells.关键词
FAT10蛋白质/真核翻译延伸因子1α1/RNA干扰Key words
FAT10 prolein/ Eukaryolic translation elongation faclor 1 alpha 1/ RNA interference分类
医药卫生引用本文复制引用
余新,刘天德,袁荣发,王庆诺,李国惠,邵江华..酵母双杂交技术鉴定FAT10新的结合蛋白:eEF1A1[J].中国病理生理杂志,2012,28(5):870-877,8.基金项目
国家自然科学基金资助项目(No.81060196) （No.81060196）
教育部科学技术研究重点项目(No.208070) （No.208070）
江西省教育厅科学技术研究重点项目(No.GJJ08003) （No.GJJ08003）
南昌大学研究生创新基金资助项目(No.YC09A015) （No.YC09A015）
