Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9126917 | Gene | 2005 | 7 Pages |
Abstract
The 79 kD gonadotropin-regulated testicular long chain acyl-CoA synthetase gene (GR-LACS) is a hormone-regulated member of the acyl-CoA synthetase family that is expressed abundantly in Leydig cells and to a lesser extent in germinal cells of the adult testis. GR-LACS possesses an ATP/AMP binding domain and the fatty acyl-CoA synthetase (FACS) signature motif. To gain insights into the transcriptional regulation of GR-LACS in gonadal cells, we determined the genomic organization of the gene, including the upstream flanking sequences. The mouse GR-LACS gene spans over at least 45 kb and the coding region is encoded by exons 1-14. All exon-intron junction sites correspond to the consensus splice sequence GT-AG. Exon 7 and 11 comprise the conserved ATP/AMP binding domain and the FACS signature motif, respectively. Primer extension and S1 nuclease analyses demonstrated four transcriptional start sites located at â 266 / â 216 bp 5â² to the ATG codon. The minimal promoter domain resides within â 254 / â 217 bp 5â² to ATG codon, and upstream sequences to â 404 bp (â 1035 / â 405 bp) contribute to the inhibition of transcription in the expressing mouse Leydig tumor cells. Removal of â 217 / â 1 bp, containing a 23 nt GC rich sequence (â 112 / â 90) with an Sp1/Sp3 binding element, within the 1st exon of this TATA-less promoter, significantly reduced GR-LACS gene transcription. Transcriptional activity was abolished by a 2 nt mutation of this element. Thus, functional analyses of this promoter domain indicate that transcription of GR-LACS gene requires an Sp1/Sp3 binding element downstream of the transcriptional start sites which is essential for basal promoter activity.
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Biochemistry, Genetics and Molecular Biology
Genetics
Authors
Yi Sheng, Jie Li, Maria L. Dufau, Chon-Hwa Tsai-Morris,