转录因子Elk-1调控MAZ基因启动子转录活性的体外研究  

Down-Regulation of Human MAZ Gene Promoter Activity by Transcription Factor Elk-1 In Vitro

作　　者：王博[1] 贾琳娜[1] 王小怡[1] 张潮[1] 王雪伟[1] 任剑波[1] 高哲[1] 郭大玮[1] Wang Bo Jia Linna Wang Xiaoyi Zhang Chao Wang Xuewei Ren Jianbo Gao Zhe Guo Dawei(Department of Forensic Science ＆ Translational Medicine Research Center, Shanxi Medical University, Taiyuan 030001, China)

机构地区：[1]山西医科大学法医学院,山西医科大学转化医学研究中心,太原030001

出　　处：《中国细胞生物学学报》2017年第6期710-719,共10页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:90608008);高等学校博士学科点专项科研基金博导类(批准号:20111417110003)资助的课题~~

摘　　要：该文旨在探究血清淀粉样蛋白A激活转录因子(serum amyloid A activating transcription factor,SAF),或称myc相关锌指蛋白(myc-associated zinc finger protein,MAZ)基因,除已证实的转录因子MAZ和Sp1(specificity protein 1)外,是否存在其他转录因子对MAZ基因启动子的转录激活具有调控作用。在生物信息学预测基础上,利用凝胶电泳迁移实验(electrophoretic mobility shift assay,EMSA)筛查MAZ启动子区的转录因子结合位点。在EMSA筛查出MAZ启动子区包含转录激活因子ETS样蛋白1(ETS-like 1 transcription factor,Elk-1)结合位点后,通过该实验室构建的由MAZ启动子驱动的双荧光报告系统分析观察过表达转录因子Elk-1后MAZ启动子的转录激活情况。结果显示,转录因子Elk-1可结合MAZ启动子–525~–504 nt区的DNA序列;过表达Elk-1及Sp1均可使MAZ的两个转录子SAF-1和SAF-3转录水平降低且后一转录子被抑制尤甚。结果提示,Elk-1与Sp1共同参与调控MAZ基因两个转录子SAF-1和SAF-3的转录调控,在MAZ基因参与的细胞诸多生理疾病过程中,Elk-1可能通过该途径发挥其作用。The aim of this work is to investigate the transcriptional regulation of the myc-associated zinc finger protein（MAZ）, also known as serum amyliod A activating transcription factor（SAF）, which plays a key role in regulation of variety of inflammation-responsive genes as well as transcription of multiple genes modulating tumor microenvironment. The down-regulation of MAZ promoter activity by Sp1 and MAZ were identified. In this study, the transcription factor ETS-like 1 transcription factor（Elk-1） binding site for regulating MAZ transcription was identified by PROMO software online and electrophoretic mobility shift assay（EMSA）. The expressions of both SAF-1 and SAF-3, the two transcription variants of MAZ gene, were analyzed by MAZ promoter driving bichromatic fluorescent reporter system when Elk-1 was over-expressed. The results showed that the region between –525 and –504 bp in MAZ promoter region could be bound with Elk-1. The expressions of both SAF-1 and SAF-3 were inhibited when Elk-1 was over-expressed. In comparison to transcription factor Sp1, it seems that Elk-1 is moderate inhibitor for MAZ promoter activation. Based on our observations, it is suggested that as a downregulation transcription factor for MAZ promoter activation, Elk-1 may play roles in physiological and pathological procedures in human cells, such as inflammation reaction and tumorigenesis. The hypothesis needs to be confirmed by further analysis in vivo.

关 键 词：MAZ基因 转录因子 Elk-1 凝胶电泳迁移实验 双色荧光蛋白报告系统 

分 类 号：Q78[生物学—分子生物学]