Characterization of the human platelet endothelial cell adhesion molecule-1 promoter: Identification of a GATA-2 binding element required for optimal transcriptional activity
Characterization of the human platelet endothelial cell adhesion molecule-1 promoter: Identification of a GATA-2 binding element required for optimal transcriptional activity
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DOI：
10.1182/blood.v89.4.1260
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发表时间：
1997-02-15
期刊：
影响因子：
20.3
通讯作者：
Newman, PJ
中科院分区：
文献类型：
--
作者：
Gumina, RJ;Kirschbaum, NE;Newman, PJ
血小板/内皮细胞粘附分子-1（PECAM-1）是IG基因超家族的130 kD成员，表达于血小板、内皮细胞和某些白细胞亚群上。为了研究控制PECAM-1血管特异性表达的因素，我们克隆了PECAM-1基因的5 '侧翼区并分析了其转录活性。5 ′-RACE分析表明，转录起始发生在翻译起始位点上游约204 bp处的几个紧密间隔的邻近位点。对转录起始位点（TIS）上游序列的分析显示没有典型的TATA或CAAT元件，然而在无TATA启动子中常见的起始元件包含TIS。克隆在荧光素酶报告基因前面的5 '-连续截短的PECAM-1启动子片段以谱系特异性和方向特异性方式驱动转录。存在于300-bp核心启动子内的推定的顺式作用控制元件包括两个ets位点、一个Spl位点、串联的E-box结构域、两个GATA相关位点（CACCC）、一个AP-2结合位点和一个位于-24的加塔元件。突变分析表明，最佳转录活性需要位置-24的加塔序列，并且凝胶迁移分析进一步表明，加塔-2转录因子，而不是加塔-1，与PECAM-1启动子的该区域结合，了解调节PECAM-1组织特异性表达的顺式和反式作用因子，将增加我们对血管内皮细胞凋亡机制的理解。实现特异性基因表达。(C)1997年，美国血液学会。
Platelet/endothelial cell adhesion molecule-1 (PECAM-1) is a 130-kD member of the Ig gene superfamily that is expressed on platelets, endothelial cells, and certain leukocyte subsets. To examine the factors controlling vascular-specific expression of PECAM-1, we cloned the 5'-flanking region of the PECAM-1 gene and analyzed its transcriptional activity. 5'-Rapid amplification of cDNA ends (5'-RACE) analysis showed that transcription initiation occurred at several closely spaced nearby sites originating approximately 204 bp upstream from the translation start site. Analysis of the sequence immediately upstream from the transcription initiation site (TIS) showed no canonical TATA or CAAT elements, however an initiator element commonly found in TATA-less promoters encompassed the TIS. 5'-serially truncated PECAM-1 promoter segments cloned in front of a luciferase reporter drove transcription in both a lineage- and orientation-specific manner. Putative cis-acting control elements present within a 300-bp core promoter included two ets sites, an Spl site, tandem E-box domains, two GATA-associated sites (CACCC), an AP-2 binding site, and a GATA element at -24. Mutational analysis showed that optimal transcriptional activity required the GATA sequence at position -24, and gel-shift assays further showed that the GATA-2 transcription factor, but not GATA-1, bound to this region of the PECAM-1 promoter, Understanding the cis- and transacting factors that regulate the tissue-specific expression of PECAM-1 should increase our understanding of the mechanisms by which vascular-specific gene expression is achieved. (C) 1997 by The American Society of Hematology.