Differential roles for ETS, CREB, and EGR binding sites in mediating VEGF receptor 1 expression in vivo
Differential roles for ETS, CREB, and EGR binding sites in mediating VEGF receptor 1 expression in vivo
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DOI：
10.1182/blood-2009-05-220434
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发表时间：
2009-12-24
期刊：
影响因子：
20.3
通讯作者：
Aird, William C.
中科院分区：
文献类型：
--
作者：
Jin, Enjing;Liu, Ju;Aird, William C.
血管内皮生长因子受体1 （VEGFR1）是内皮特异性基因表达的标志物。我们之前报道了人类VEGFR1启动子（-748和+284之间）包含在转基因小鼠完整内皮中表达的信息。本研究的目的是解剖体外和体内VEGFR1启动子活性的顺式调控元件。在原代内皮细胞中，e74样因子1 （ELF-1，介于-49和-52之间）、环腺苷单磷酸反应元件结合位点（CREB，介于- 74和- 81之间）和早期生长反应因子1/3 （EGR-1/3，介于- 16和- 25之间）的结合位点被证明在基因转录中发挥积极作用，而假设的E26转化特异性序列（ETS）基序，介于- 36和- 39之间，对启动子活性有净负面影响。当靶向小鼠的Hprt位点时，ELF-1结合位点和CRE元件的突变降低了胚胎血管中启动子的活性，并导致成人内皮中启动子的表达缺失。出生后，EGR结合位点突变体在一个血管床亚群中表现出显著降低的启动子活性。相反，- 39 ETS位点的突变导致多个血管床LacZ染色增加。总之，这些结果为VEGFR1的转录调控机制提供了新的见解。[血液。2009;114:5557-5566]
Vascular endothelial growth factor receptor 1 (VEGFR1) is a marker for endothelial-specific gene expression. We previously reported that the human VEGFR1 promoter (between -748 and +284) contains information for expression in the intact endothelium of transgenic mice. The objective of this study was to dissect the cis-regulatory elements underlying VEGFR1 promoter activity in vitro and in vivo. In primary endothelial cells, binding sites for E74-like factor 1 (ELF-1; between -49 and -52), cyclic adenosine monophosphate response element binding (CREB; between - 74 and - 81), and early growth response factor 1/3 (EGR-1/3; between - 16 to - 25) were shown to play a positive role in gene transcription, whereas a putative E26 transformation-specific sequence (ETS) motif between - 36 and - 39 had a net negative effect on promoter activity. When targeted to the Hprt locus of mice, mutations of the ELF-1 binding site and the CRE element reduced promoter activity in the embryonic vasculature and resulted in a virtual loss of expression in adult endothelium. Postnatally, the EGR binding site mutant displayed significantly reduced promoter activity in a subset of vascular beds. In contrast, mutation of the - 39 ETS site resulted in increased LacZ staining in multiple vascular beds. Together, these results provide new insights into the transcriptional regulatory mechanisms of VEGFR1. (Blood. 2009; 114: 5557-5566)