人细胞色素P-45011β的cDNA和基因组DNA的克隆

Cloning of cDNA and genomic DNA for human cytochrome P-45011 beta.

作者信息

Kawamoto T, Mitsuuchi Y, Toda K, Miyahara K, Yokoyama Y, Nakao K, Hosoda K, Yamamoto Y, Imura H, Shizuta Y

机构信息

Department of Medical Chemistry, Kochi Medical School, Japan.

出版信息

FEBS Lett. 1990 Sep 3;269(2):345-9. doi: 10.1016/0014-5793(90)81190-y.

DOI:10.1016/0014-5793(90)81190-y

PMID:2401360

Abstract

A full-length cDNA clone encoding steroid 11 beta-hydroxylase (P-45011 beta) has been isolated from a cDNA library derived from human adrenal tumor. The insert of the clone contains an open reading frame encoding a protein of 503 amino acid residues together with a 4 bp 5'-untranslated region and a 576 bp 3'-untranslated region to which a poly(A) tract is attached. The promoter region of the P-450(11) beta gene has also been isolated from a genomic library derived from human pre-B cells. It contains a TATA box, a putative cAMP-responsive element, several repeated sequences and two sequence elements similar to the consensus sequence for binding of AP-1. A transient expression assay in Y-1 adrenal tumor cells demonstrates that the promoter activity is remarkably enhanced by treatment of the cells with cAMP. In addition, analysis using deletion mutants containing various lengths of the 5'-flanking region of the gene suggests that several cis-acting elements participate in transcriptional regulation of human P-450(11) beta gene.

摘要

已从源自人肾上腺肿瘤的cDNA文库中分离出编码类固醇11β-羟化酶（P-45011β）的全长cDNA克隆。该克隆的插入片段包含一个编码503个氨基酸残基蛋白质的开放阅读框，以及一个4bp的5'-非翻译区和一个576bp的3'-非翻译区，3'-非翻译区连接有一个聚腺苷酸尾。P-450(11)β基因的启动子区域也已从源自人前B细胞的基因组文库中分离出来。它包含一个TATA盒、一个假定的cAMP反应元件、几个重复序列以及两个与AP-1结合共有序列相似的序列元件。在Y-1肾上腺肿瘤细胞中的瞬时表达分析表明，用cAMP处理细胞可显著增强启动子活性。此外，使用含有该基因不同长度5'-侧翼区域的缺失突变体进行的分析表明，几个顺式作用元件参与了人P-450(11)β基因的转录调控。