Regulation of transcription of the human presenilin-1 gene by Ets transcription factors and the p53 protooncogene
Regulation of transcription of the human presenilin-1 gene by Ets transcription factors and the p53 protooncogene
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DOI：
10.1074/jbc.m005411200
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发表时间：
2000-11-10
影响因子：
4.8
通讯作者：
Das, HK
中科院分区：
文献类型：
--
作者：
Pastorcic, M;Das, HK
人早老素-1细胞基因的表达被p53原癌基因抑制。下调的快速动力学表明，它可能是由一个初级机制引起的。我们在这里表明，p53也抑制转录的早老素-1启动子氯霉素乙酰转移酶报告合成基因在神经母细胞瘤（SK-N-SH）和肝癌（HepG 2）细胞系的瞬时感染试验。仅：包括从转录起始起-35 td + 6的序列的最小启动子足以赋予下调。我们以前已经定义了一个关键的DNA元件控制90%的基因表达：在同一个短区域内，并确定所涉及的转录因子也应该提供的PSI的调节p53的见解。该区域包含Ets转录因子结合基序，并且Ets共有序列的核心序列内的a-碱基对改变（GGAA至TTAA）也使转录降低超过90%。我们现在发现，Ets 1和Ets 2：确实反式激活PSI启动子-氯霉素乙酰转移酶报告，包括（-35至+6）：片段。此外，在电泳迁移率变动测定中，体外翻译的Ets 2特异性结合至-10 Ets基序。因此，Ets 1/2因子特异性结合到-10 Ets元件并激活PS1转录。我们还表明，辅激活因子p300增强激活Ets 1和Ets 2以及抑制p53。已知p300与p53以及Ets 1和Ets 2相互作用。我们发现，p53不直接结合PSI启动子。因此，P53对PS1转录的抑制可能是通过蛋白质-蛋白质相互作用介导的。
The expression of the human presenilin-1 cellular gene is suppressed by the p53 protooncogene. The rapid kinetic of the down-regulation has suggested that it may result from a primary mechanism. We show here that p53 also suppresses the transcription of a presenilin-1 promoter-chloramphenicol acetyltransferase reporter synthetic gene in transient infection assays in neuroblastoma (SK-N-SH) and hepatoma (HepG2) cell lines. Only:a minimum promoter including sequences from -35 td + 6 from the transcription initiation is sufficient to confer down-regulation. We have previously defined a crucial DNA element controlling 90% of the expression of the gene:within the same short area, and the identification of the transcription factors involved should also provide insights into the regulation of PSI by p53. This region,contains an Ets transcription factor binding motif, and a a-base pair alteration within the core sequence (GGAA to TTAA) of the Ets consensus also reduced transcription by more than 90%. We now show that Ets1 and Ets2: indeed transactivate a PSI promoter-chloramphenicol acetyltransferase reporter including the (-35 to +6):fragment. Furthermore, in vitro translated Ets2 binds specifically to the -10 Ets motif in electrophoretic mobility shift assays. Therefore, Ets1/2 factors bind specifically to the -10 Ets element and activate PS1 transcription. We also show that the coactivator p300 enhances the activation by Ets1 and Ets2 as well as the repression by p53. p300 is known to interact with p53 as well as with Ets1 and Ets2. We show that p53 does not bind directly to the PSI promoter. Hence the repression of PS1:transcription by p53 is likely to be mediated through protein-protein interactions.