The mammalian alcohol dehydrogenases (ADHs) are a gene family whose products catalyze oxidation of alcohol to corresponding aldehyde, itaticizeADHis a relatively newly discovered member of the human ADH gene family; its$function and regulation have not been well studied. In order to test the promoter activity of this gene, several different lengths of the italicizeADH6 promoter were inserted into pXP2, a vector that has a promoterless luciferase reporter gene. All constructs tested showed a much stronger activity than pXP2 alone (10 tol00-fold increase) in two cell lines:CV-1 and H4IIE-C3. This demonstrates that the italicizeADH6 promoter is functional in these cell lines. Similar patterns of promoter activity were observed in both cell lines, suggesting that the major determinants of tissue-specificity may be missing in these short promoter fragments. Therefore, a human genomic DNA library was screened and clones containing the upstream region of the italicizeADH6 promoter were obtained. Three longer promoter fragments (0.55 kb,l.6 kb and 2.3 kb) were cloned into pXP2 and tested for promoter activity. Different patterns of activity were observed in two cell lines, suggesting the existence of tissuespecific elements in the upstream region of the italicizeADH6promoter. DNaseI footprinting assays show several sites in the proximal region were protected by proteins present in liver extract. One of these sites interacts with C/EBP. Mutation of this site drastically decreases the promoter activity, suggesting this C/EBP site plays an important role in italicizeADH6 regulation. [Supported by N.I.A.A.A. R01-AA06460.
|Original language||English (US)|
|State||Published - Dec 1 1997|
ASJC Scopus subject areas
- Molecular Biology