Identification of regulatory sequences in the type 1 plasminogen activator inhibitor gene responsive to transforming growth factor beta.
Identification of regulatory sequences in the type 1 plasminogen activator inhibitor gene responsive to transforming growth factor beta.
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DOI：
10.1016/s0021-9258(18)54461-6
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发表时间：
1991-12
期刊：
影响因子：
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通讯作者：
M. Keeton;S. Curriden;A. J. Zonneveld;D. Loskutoff
中科院分区：
文献类型：
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作者：
M. Keeton;S. Curriden;A. J. Zonneveld;D. Loskutoff
研究了转化生长因子β（TGF β）对人1型纤溶酶原激活物抑制剂（派-1）启动子的调节。将来自派-1启动子和5 '侧翼区的800个碱基对片段与萤火虫荧光素酶报告基因融合，并转染到Hep 3B人肝癌细胞中。用TGF β处理细胞诱导荧光素酶活性超过50倍。使用通过该区域具有5'或3'缺失的构建体的转染研究揭示了两个序列在TGF β应答中是重要的。第一个序列位于近端启动子（-49至-87），并介导TGF β的11倍诱导，而第二个更远端的区域（-636至-740）包含两个序列，它们一起介导50倍或更大的应答。序列比较表明，这两个响应区域含有与AP-1的共有结合位点具有高度同源性的序列。此外，凝胶阻滞分析实验表明，这两个序列结合一个共同的核蛋白，并且含有一个共识AP-1序列的寡核苷酸能够竞争这种共同的蛋白质的结合。因此，派-1基因对TGF β的应答由至少两个独立的区域介导，并且这两个区域都含有与AP-1结合位点同源的DNA序列。
Regulation of the human type 1 plasminogen activator inhibitor (PAI-1) promoter by transforming growth factor-beta (TGF beta) was studied. An 800-base pair fragment from the PAI-1 promoter and 5'-flanking region was fused to the firefly luciferase reporter gene and transfected into Hep3B human hepatoma cells. Treatment of the cells with TGF beta induced luciferase activity by more than 50-fold. Transfection studies using constructs with 5' or 3' deletions through this region revealed that two sequences were important in the TGF beta response. The first sequence was located in the proximal promoter (-49 to -87) and mediated an 11-fold induction with TGF beta, while the second more distal region (-636 to -740) contained two sequences which together mediated a 50-fold or greater response. Sequence comparison indicated that both of the responsive regions contained sequences with high homology to the AP-1 consensus binding site. Moreover, gel retardation analysis experiments demonstrated that both sequences bound a common nuclear protein, and that an oligonucleotide containing a consensus AP-1 sequence was able to compete for the binding of this common protein. Thus, the response of the PAI-1 gene to TGF beta is mediated by at least two separate regions, and both of these regions contain DNA sequences homologous to the AP-1 binding site.