Characterization of the regulatory element(s) controlling the adrenal-specific expression of human CYP21(P450c21) gene.
| Autor: | Cheng Chai-Li, 鄭佳俐 |
|---|---|
| Rok vydání: | 1998 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 86 Steroid 21 hydroxylase (P450c21) is specifically expressed in the adrenal cort ex, and is required for the biosynthesis of steroid hormones. Enzymatic defici ency of 21-hydroxylase causes the congenital adrenal hyperplasia (CAH), an aut osomal recessive disorder. There are two copies of human steroid 21-hydroxylas e gene, the active CYP21 which encodes the enzyme, and the pseudogene, CYP21P. These two genes are localized on human chromosome 6p21.1, and duplicated in t andem with the genes for the fourth components of complement, C4A and C4B, and other newly characterized duplicated genes, organized as following: 5''C4A(ZA( CYP21P(YA(XA(C4B(ZB(CYP21(YB(XB-S3''. In this complex locus, most of the transc ripts are adrenal-specific, including the CYP21, XA, YA, ZA,. XB-S, YB, ZB.To study the cis-elements controlling the adrenal-specific expression of these ge nes, we analyzed the effects of DNA sequences upstream from the CYP21P and CYP 21 on the transcription activity of the basic promoter of the CYP21. We used m ouse adrenocorticoid tumor Y-1 cell, testicular Leydig MA10 cell, and human li ver Hep G-2 tumor cells to analyze the adrenal-specific expression. After tran sient transfection into cells, the transcription activity was analyzed using C AT and primer extension assays. The results showed that DNA sequences at the - 6328/-1668 bp region upstream from the CYP21 enhanced the transcriptional acti vity of CYP21 to 2 fold in Y-1 cell, but not in MA10 or Hep G-2 cells. However, smaller DNA fragments covering the same region didn''t show enhanced effect but suppressed the transcription activity of the CYP21 in all three cell lines tested. In addition, DNA regions within the 13.5 kb upstream from the CYP21P all suppressed the transcription activity of the CYP21 but to a different leve l. Therefore, we predict the regulatory element(s) controlling the adrenal-spe cific expression of the CYP21 may be located within the -6328/-1668 bp region. Other cis-elements at the C4/CYP21 gene locus which are not tested in this st udy may also be involved in the regulation of the tissue-specific expression o f human steroid 21-hydroxylase. Further study will be performed to test this p ossibili |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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