In an effort to search for transcription factors involved in ADH gene expression, we have examined the effects of multimers of individual cis-arting elements inserted in front of an SV40 early promoter in the iuciferase expression vector pGL2-Promoter (Promega). A trimer of a ubiquitous site in ADH3 (for which we had not identified a known transcription factor) showed weak enhancer-like activity, increasing transcription 1.5- to 2-fold regardless of tho orientation in which it was inserted. Using this trimer as ligand to screen a human liver cDNA expression library, we identified a positive clone which appears to be a novel transcription factor. Competition studies suggests that the binding to the ligand is sequence specific. We rescreened the human liver cDNA library and identified nine overlapping clones representing 3.-4 kb of cDNA. The nucleotide sequence of the coding region has extremely high G-I-C content (75.5%). This cDNA is not closely related to any sequence in the GenBank database, except for a region that contains Iwo tandem C2 H2 zinc fingers that match the consensus well. The conserved zinc finger motif probably serves as a DNA binding domain. The deduced amino acid sequence of this putative transcription factor contains a very hydrophilic region, and ends with a cluster of stop signals. [Supported by N.I.A.A.A. R01-AA06460.].
|Original language||English (US)|
|State||Published - Dec 1 1996|
ASJC Scopus subject areas
- Molecular Biology