Manohar A, Anwar R A
Department of Biochemistry, University of Toronto, Ontario, Canada.
Biochem J. 1994 May 15;300 ( Pt 1)(Pt 1):147-52. doi: 10.1042/bj3000147.
A cell-specific negative regulatory element has been identified in the first intron of the gene for elastin in a region between 442 and 464 bp from the translational start site. This regulatory element functions both when it is located 5' of the promoter and 3' of the chloramphenicol acetyltransferase (CAT) gene. The inhibition is observed both with the homologous elastin promoter and the heterologous SV1 promoter in transient expression experiments using rat aortic smooth-muscle cells. No inhibition was observed with NIH 3T3, Hep G2 and little, if any, with HeLa cells. Cell specificity was further confirmed by DNA mobility shift assays and the position of the negative regulatory element was localized with the use of synthetic duplex oligomers. It is proposed that this negative element plays a significant role in the modulation of the expression of the gene for elastin in the smooth-muscle cells of the aorta during development.
在弹性蛋白基因的第一个内含子中,已在距翻译起始位点442至464 bp的区域内鉴定出一种细胞特异性负调控元件。当该调控元件位于启动子的5'端和氯霉素乙酰转移酶(CAT)基因的3'端时均发挥作用。在使用大鼠主动脉平滑肌细胞的瞬时表达实验中,同源弹性蛋白启动子和异源SV1启动子均观察到抑制作用。在NIH 3T3细胞、Hep G2细胞中未观察到抑制作用,在HeLa细胞中即使有抑制作用也很微弱。通过DNA迁移率变动分析进一步证实了细胞特异性,并使用合成双链寡聚物确定了负调控元件的位置。有人提出,这种负调控元件在发育过程中对主动脉平滑肌细胞中弹性蛋白基因表达的调节起着重要作用。