Inoue A, Yanagisawa M, Takuwa Y, Mitsui Y, Kobayashi M, Masaki T
Institute of Basic Medical Sciences, University of Tsukuba, Ibaraki, Japan.
J Biol Chem. 1989 Sep 5;264(25):14954-9.
Endothelin-1 is a 21-amino acid potent vasoconstrictor peptide produced by vascular endothelial cells. We have cloned the whole length of the human preproendothelin-1 (PPET-1) gene and the corresponding cDNA and determined the complete nucleotide sequences. The 2026-nucleotide human mRNA for PPET-1 (excluding the polY(A) tail) is encoded in five exons distributed over 6836 base pairs of the genome. The 5'-flanking region of the gene contains (i) octanucleotide sequences for the phorbol ester-responsive elements, also known as the binding elements for FOS.JUN complex; (ii) consensus motifs for the binding site of nuclear factor 1, which may mediate the induction described previously of PPET-1 mRNA by transforming growth factor-beta; (iii) hexanucleotide sequences for the acute phase reactant regulatory elements that may be involved in the induction of endothelin-1 under acute physical stress in vivo. Further, the 3'-nontranslated sequence of human PPET-1 mRNA contains three AUUUA motifs, which may mediate selective translation-dependent destabilization of the mRNA. Northern blot analysis in cultured endothelial cells from human umbilical veins shows that PPET-1 mRNA is in fact rapidly induced by the active phorbol ester 12-O-tetradecanoylphorbol 13-acetate within 10 min. Analysis of mRNA life span by using actinomycin D demonstrates that PPET-1 mRNA has a short intracellular half-life of about 15 min and is superinduced by cycloheximide. This superinduction is found to be due to the stabilization of the mRNA by cycloheximide, as in the case of other known AUUUA-containing mRNAs. These findings suggest that the regulation of expression of PPET-1 mRNA may be mediated in part by these sequence elements.
内皮素-1是一种由血管内皮细胞产生的含21个氨基酸的强效血管收缩肽。我们已经克隆了人前内皮素-1（PPET-1）基因的全长及相应的cDNA，并确定了完整的核苷酸序列。PPET-1的2026个核苷酸的人mRNA（不包括聚腺苷酸尾）由分布在基因组6836个碱基对中的五个外显子编码。该基因的5'侧翼区域包含：（i）佛波酯反应元件的八核苷酸序列，也称为FOS-JUN复合物的结合元件；（ii）核因子1结合位点的共有基序，其可能介导先前描述的转化生长因子-β对PPET-1 mRNA的诱导；（iii）急性期反应物调节元件的六核苷酸序列，其可能参与体内急性身体应激下内皮素-1的诱导。此外，人PPET-1 mRNA的3'非翻译序列包含三个AUUUA基序，其可能介导mRNA的选择性翻译依赖性去稳定化。对人脐静脉培养的内皮细胞进行的Northern印迹分析表明，活性佛波酯12-O-十四酰佛波醇13-乙酸酯实际上在10分钟内迅速诱导了PPET-1 mRNA。使用放线菌素D分析mRNA寿命表明，PPET-1 mRNA在细胞内的半衰期约为15分钟，并且被环己酰亚胺超诱导。发现这种超诱导是由于环己酰亚胺使mRNA稳定，如同其他已知的含AUUUA的mRNA的情况一样。这些发现表明，PPET-1 mRNA表达的调节可能部分由这些序列元件介导。
