Kosaka T, Miyata A, Ihara H, Hara S, Sugimoto T, Takeda O, Takahashi E, Tanabe T
Department of Pharmacology, National Cardiovascular Center Research Institute, Osaka, Japan.
Eur J Biochem. 1994 May 1;221(3):889-97. doi: 10.1111/j.1432-1033.1994.tb18804.x.
The human gene (PTGS2) encoding an inducible isozyme of prostaglandin-endoperoxide synthase (prostaglandin-endoperoxide synthase 2) that is distinct from the well-characterized and constitutive isozyme (prostaglandin-endoperoxide synthase 1), was isolated using a polymerase-chain reaction-generated cDNA fragment probe for human prostaglandin-endoperoxide synthase 2. Nucleotide sequence analysis of the entire human prostaglandin-endoperoxide-synthase-2 gene demonstrated that it is more than 8.3 kb in size and consists of ten exons; this gene is very similar to the murine and chicken prostaglandin-endoperoxide-synthase-2 genes. The structures of exons in the human prostaglandin-endoperoxide-synthase-2 gene were also similar to those of the human prostaglandin-endoperoxide-synthase-1 gene (PTGS1). However, the sizes of introns in the human prostaglandin-endoperoxide-synthase-2 gene were generally smaller than those of the human prostaglandin-endoperoxide-synthase-1 gene. Primer-extension analysis indicated that the transcriptional-start site is 134 bases upstream of the translational-initiation site. The sequence of the 1.69-kb region of nucleotides preceding the transcriptional-start site and the first 0.8-kb intron contained a canonical TATA box and various transcriptional-regulatory elements (CArG box, NF-IL6, PEA-1, myb, GATA-1, xenobiotic-response element, cAMP-response element, NF-kappa B, PEA-3, Sp-1 and 12-O-tetradecanoyl-phorbol-13-acetate-response element). The nucleotide sequence of the 5'-flanking region (275 bp) of the human prostaglandin-endoperoxide-synthase-2 gene showed 63% similarity to the sequence of murine prostaglandin-endoperoxide-synthase-2/TIS10 gene, but essentially no homology to the chicken prostaglandin-endoperoxide-synthase-2 gene, and human and murine prostaglandin-endoperoxide-synthase-1 genes. A fluorescence in situ hybridization study showed that the human genes coding for prostaglandin-endoperoxide synthase 1 (PTGS1) and prostaglandin-endoperoxidase synthase 2 (PTGS2) were mapped to distinct chromosomes 9q32-q33.3 and 1q25.2-q25.3, respectively, indicating that these genes are not genetically linked.
编码前列腺素内过氧化物合酶诱导型同工酶（前列腺素内过氧化物合酶2）的人类基因（PTGS2）与已充分表征的组成型同工酶（前列腺素内过氧化物合酶1）不同，使用针对人类前列腺素内过氧化物合酶2的聚合酶链反应生成的cDNA片段探针进行分离。对整个人类前列腺素内过氧化物合酶2基因的核苷酸序列分析表明，其大小超过8.3 kb，由十个外显子组成；该基因与小鼠和鸡的前列腺素内过氧化物合酶2基因非常相似。人类前列腺素内过氧化物合酶2基因中外显子的结构也与人类前列腺素内过氧化物合酶1基因（PTGS1）的相似。然而，人类前列腺素内过氧化物合酶2基因中内含子的大小通常比人类前列腺素内过氧化物合酶1基因的小。引物延伸分析表明转录起始位点在翻译起始位点上游134个碱基处。转录起始位点之前1.69 kb的核苷酸区域和第一个0.8 kb内含子的序列包含一个典型的TATA盒和各种转录调控元件（CArG盒、NF-IL6、PEA-1、myb、GATA-1、外源性反应元件、cAMP反应元件、NF-κB、PEA-3、Sp-1和12-O-十四烷酰佛波醇-13-乙酸反应元件）。人类前列腺素内过氧化物合酶2基因5'侧翼区域（275 bp）的核苷酸序列与小鼠前列腺素内过氧化物合酶2/TIS10基因的序列显示63%的相似性，但与鸡前列腺素内过氧化物合酶2基因以及人类和小鼠前列腺素内过氧化物合酶1基因基本没有同源性。荧光原位杂交研究表明，编码前列腺素内过氧化物合酶1（PTGS1）和前列腺素内过氧化物合酶2（PTGS2）的人类基因分别定位于不同的染色体9q32 - q33.3和1q25.2 - q25.3，表明这些基因没有遗传连锁关系。
