转录因子MAZ对c1orf109基因转录表达调控的体外研究

Regulation of c1orf109 gene by transcription factor MAZ in vitro

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He La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。

AIM:To study the regulation of human chromosome1open reading frame109(c1orf109)gene by transcription factor Myc-associated zinc-finger protein(MAZ)in vitro.METHODS:In vitro study,electrophoretic mobility shift assay was performed to screen the binding sites of MAZ in the promoter region of c1orf109gene.The HeLa cells were co-transfected with the enhanced green fluorescent protein reporter vector driven by c1orf109promoter,and MAZ and transcription factor specificity protein1(Sp1)expression plasmids.After24h,the transcriptional expression of c1orf109gene in the co-transfected cells was determined by confocal scanning microscopy and flow cytometry.RESULTS:The c1orf109promoter could bind to MAZ,and shared the binding sites with Sp1.MAZ and Sp1both inhibited the transcriptional expression of c1orf109gene and the inhibition effect of Sp1was greater than MAZ(P<0.05).CONCLUSION:Both MAZ and Sp1regulate the expression of c1orf109gene 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 c1orf109gene is vital important for cell biological processing.