Timmers H T, Pronk G J, Bos J L, van der Eb A J
Laboratory for Molecular Carcinogenesis, Sylvius Laboratories, University of Leiden, The Netherlands.
Nucleic Acids Res. 1990 Jan 11;18(1):23-34. doi: 10.1093/nar/18.1.23.
We have cloned the immediate-early serum-reponsive JE gene from the rat in order to study the regulation of this gene. We show that sequences of the JE promoter region confer serum-inducibility to a reporter gene. Analysis of the promoter in transient assays reveals that: i) the -141/-88 region is required for the response to the phorbol ester TPA, ii) the -70/-38 region is essential for basal activity. This latter region harbors the sequence TGACTCC, which resembles the consensus site for AP-1 binding, TGACTCA. DNA-protein binding assays indicate that the JE AP-1 site and the consensus AP-1 site have an overlapping but not identical binding spectrum for AP-1 proteins. Our data suggest that the inability of some AP-1 sites to respond to TPA is caused by subtle differences in affinity for AP-1 proteins.
为了研究该基因的调控机制,我们从大鼠中克隆了早期血清反应性JE基因。我们发现JE启动子区域的序列可赋予报告基因血清诱导性。在瞬时分析中对启动子的分析表明:i) -141 / -88区域是对佛波酯TPA产生反应所必需的,ii) -70 / -38区域对基础活性至关重要。后一个区域含有序列TGACTCC,它类似于AP-1结合的共有位点TGACTCA。DNA-蛋白质结合试验表明,JE AP-1位点和共有AP-1位点对AP-1蛋白具有重叠但不完全相同的结合谱。我们的数据表明,某些AP-1位点无法对TPA作出反应是由于对AP-1蛋白的亲和力存在细微差异所致。