转录因子MAZ对c1orf109基因转录表达调控的体外研究

摘要

目的:本文旨在研究转录因子Myc相关锌指蛋白(MAZ)在体外对人类1号染色体开放阅读框109(c1orf109)基因的转录表达调控.方法:体外条件下,采用凝胶电泳迁移实验筛选c1orf109基因启动子区MAZ的结合位点,并利用本室构建的由c1orf109启动子驱动的增强型绿色荧光蛋白报告载体与MAZ和转录因子特化蛋白1(Sp1)的表达质粒共转染HeLa细胞,24 h后,用激光共聚焦显微镜和流式细胞术检测c1orf109基因的转录表达情况.结果:c1orf109启动子区可与MAZ结合,且有与Sp1共享的结合位点;MAZ与Sp1皆可抑制c1orf109的转录表达,且Sp1的抑制作用大于MAZ(P<0.05).结论:MAZ与Sp1两个转录因子共同调控c1orf109基因在生理及病理条件的表达,且调控方向一致.这种冗余机制调控方式的存在提示该基因的精确表达调控对于细胞行使其生物学功能可能具有重要意义%AIM:To study the regulation of human chromosome 1 open reading frame 109(c1orf109)gene by transcription factor Myc-associated zinc-finger protein(MAZ)in vitro.METHODS:In vitro study,electrophoretic mobili-ty shift assay was performed to screen the binding sites of MAZ in the promoter region of c1orf109 gene.The HeLa cells were co-transfected with the enhanced green fluorescent protein reporter vector driven by c1orf109 promoter,and MAZ and transcription factor specificity protein 1(Sp1)expression plasmids.After 24 h,the transcriptional expression of c1orf109 gene in the co-transfected cells was determined by confocal scanning microscopy and flow cytometry.RESULTS:The c1orf109 promoter could bind to MAZ ,and shared the binding sites with Sp 1.MAZ and Sp1 both inhibited the transcrip-tional expression of c1orf109 gene and the inhibition effect of Sp1 was greater than MAZ(P<0.05).CONCLUSION:Both MAZ and Sp1 regulate the expression of c1orf109 gene in physiological and pathological conditions ,and this regulation is redundancy with the same direction.The existence of redundancy transcriptional regulation manner of this gene suggests that precise regulation of c1orf109 gene is vital important for cell biological processing.