Kelavkar U, Wang S, Montero A, Murtagh J, Shah K, Badr K
Center for Glomerulonephritis, Renal Division Emory University, and Veterans Affairs Medical Center, Atlanta, GA 30033, USA.
Mol Biol Rep. 1998 Jul;25(3):173-82. doi: 10.1023/a:1006813009006.
In order to study the transcriptional control of 15-LO expression, we have cloned and sequenced the human 15-LO promoter region. The 15-LO promoter is associated with a CpG island at the 5'-end of the gene, and sequence analysis reveals putative Sp1 and Ap2 binding site/s and absence of TATA or CAAT motifs. Transcription is initiated at one major site. Using deletion constructs, we have defined an active promoter region of 1056 bp. Gel-shift assays revealed that transcriptional factor(s) induced only in response to IL-13 treatment of human peripheral blood monocytes bind to the 15-LO promoter DNA. Two regions, DP1 (-140 to -92 bp) and DP2 (-353 to -304 bp) of the promoter were essential for transcription in HeLa cells and human peripheral monocytes. Hela nuclear extracts contained a specific nuclear factor(s) binding to 15-LO promoter DNA which are distinct from those derived from IL-13-treated human peripheral monocyte nuclear extracts. In addition, fluorescent in situ hybridization (FISH) results refined the previous localization of 15-LO to human chromosome 17p13.3.
为了研究15-脂氧合酶（15-LO）表达的转录调控，我们克隆并测序了人15-LO启动子区域。15-LO启动子与该基因5'端的一个CpG岛相关，序列分析揭示了推定的Sp1和Ap2结合位点，且不存在TATA或CAAT基序。转录起始于一个主要位点。使用缺失构建体，我们确定了一个1056 bp的活性启动子区域。凝胶迁移试验表明，仅在白细胞介素-13处理人外周血单核细胞后诱导产生的转录因子与15-LO启动子DNA结合。启动子的两个区域，即DP1（-140至-92 bp）和DP2（-353至-304 bp）对于HeLa细胞和人外周单核细胞中的转录至关重要。HeLa细胞核提取物含有一种与15-LO启动子DNA结合的特异性核因子，它不同于来自白细胞介素-13处理的人外周单核细胞核提取物中的核因子。此外，荧光原位杂交（FISH）结果进一步明确了先前15-LO在人染色体17p13.3上的定位。
