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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 purpose of this study was to investigate whether serum amyloid A activating transcription factor (SAF), or myc-associated zinc finger protein (MAZ) gene, And Sp1 (specificity protein 1), whether there are other transcription factors have a regulatory role in the transcriptional activation of the MAZ gene promoter. Based on the bioinformatics prediction, electrophoretic mobility shift assay (EMSA) was used to screen the transcription factor binding sites in the MAZ promoter region. After the EMSA screened MAZ promoter region containing the Elk-1 binding site, the dual fluorescent reporter system driven by the MAZ promoter The transcriptional activation of MAZ promoter after overexpression of Elk-1 was observed and analyzed. The results showed that the transcription factor Elk-1 could bind to the DNA sequence of -525 ~ -504 nt in MAZ promoter. Overexpression of Elk-1 and Sp1 both reduced the transcription level of SAF-1 and SAF-3 in two MAZ transcripts The latter transcript is particularly suppressed. The results suggest that Elk-1 and Sp1 are involved in the regulation of transcription of SAF-1 and SAF-3, two of the MAZ genes. Through this pathway, Elk-1 may play its role in many physiological diseases involving MAZ gene .