Organization and structure of the mouse gamma-glutamyl hydrolase gene and the functional identification of its promoter.

Abstract

The organization and structure of the murine GH gene encoding gamma-glutamyl hydrolase were determined. The murine GH gene spans 24kb and was found to be distributed in nine exons. The intron/exon coding junctions delineated conformed to the GT-AG rule. The 5' UTR and 3' UTR along with some coding sequences were incorporated in exons 1 and 9, respectively, whereas exons 2-8 incorporated only coding sequence. A relatively GC-rich region of the genome 5' of exon 1 was distinctly promoter-like and encoded a number of putative cis-acting elements, including six Sp1 sites known to be involved in the regulation of transcription but no TATA sequence motif. Primer extension analysis of this region with mouse liver and S180 cell mRNA revealed several tsps within the region encompassing the Sp1 sites. Functional analysis of this 5' upstream region of sequence was carried out by inserting it into the pGL3 reporter gene vector for transfection into NIH3T3 cells. The transcription of the luciferase gene that resulted in these cells established the identity of this region as an active promoter for the mouse GH gene.