TTF-1相关蛋白26磷酸化位点对人SP-B基因表达的影响  

Effect of the Phosphorylation Sites of the Thyroid Transcription Factor-1 Associated Protein 26 on Surfactant Protein-B Expression

作　　者：柴新群[1] 邓飞涛[2] 

机构地区：[1]华中科技大学同济医学院附属协和医院肝胆外科,武汉430022 [2]华中科技大学同济医学院附属协和医院妇产科,武汉430022

出　　处：《华中科技大学学报（医学版）》2007年第6期707-710,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的探讨甲状腺转录因子-1相关蛋白26(TAP26)对人肺表面活性蛋白-B(hSP-B)基因表达的调控机制。方法采用定点诱变PCR技术,分别突变掉TAP26的4个磷酸化位点(S48,S66,T219和T167S168),并获得TAP26的4个相应突变体;通过体外细胞共转染和荧光素酶报告基因活性值检测技术分别检测上述4个突变体对hSP-B基因启动子活性的影响。结果与野生型TAP26比较,突变体质粒TAP26(T167S168→V167A168)、TAP26(S48→A48)和TAP26(T219→V219)分别与hSP-B基因启动子共转染后所测荧光素酶活性值无明显差异(P>0.05);而当DNA浓度为400 ng的突变体质粒TAP26(S66→A66)与hSP-B基因启动子共转染后所测荧光素酶活性值则明显降低(P<0.05)。结论突变体质粒TAP26(S66→A66)不能促进hSP-B的表达,从而TAP26的S66蛋白激酶C(PKC)位点是TAP26移位到细胞核的重要部位。Objective To investigate the regulatory effects of thyroid transcription factor-1 associated protein 26 （TSP26） on the expression of human surfactant protein-B （hSP-B） gene. Methods Site-directed mutagenesis PCR was employed to eliminate 4 phosphorylation sites （S^48 , S^66 , T^219 and T^167 S^168 ） of TAP26 respectively and produce 4 mutants. Co-transfection study and luciferase activity measurement were performed to assay the effects of 4 mutants on the hSP-B gene expression. Resuits Luciferase activity measurement results showed that after co-transfection of three mutants of the TAP26 （T^167S^168→V^167 A^168）, TAP26 （S^48→A^48） and TAP26 （T^219→V^219） plasmid respectively with hSP-B gene promoter, the luciferase activity had no significant changes as compared with wild type TAP26 （P〉0.05）. However, after the co-transfection of 400 ng DNA plasmid of TAP26 mutants （S^66→A^66） with hSP-B gene promoter, the luciferase activity was greatly reduced （P〉0.05）. Conclusion The mutant TAP26 （S^66→A^66） couldn＇t promote the expression of hSP-B, suggesting the S^66 PKC site is important for TAP26 translocation into the nucleus.

关 键 词：甲状腺转录因子-1相关蛋白26 肺表面活性蛋白-B 基因调节 

分 类 号：Q344.14[生物学—遗传学]