Regulation of the expression of the sn-glycerol-3-phosphate dehydrogenase gene in Drosophila melanogaster

Citation
S. Bartoszewski et Jb. Gibson, Regulation of the expression of the sn-glycerol-3-phosphate dehydrogenase gene in Drosophila melanogaster, BIOCHEM GEN, 36(9-10), 1998, pp. 329-350
Citations number
42
Categorie Soggetti
Molecular Biology & Genetics
Journal title
BIOCHEMICAL GENETICS
ISSN journal
00062928 → ACNP
Volume
36
Issue
9-10
Year of publication
1998
Pages
329 - 350
Database
ISI
SICI code
0006-2928(199810)36:9-10<329:ROTEOT>2.0.ZU;2-C
Abstract
P element-mediated transformation has been used to investigate the regulati on of expression of the sn-glycerol-3-phosphate dehydrogenase gene of Droso phila melanogaster. A 13-kb construct containing the eight exons and associ ated introns, 5 kb of the 5' region, and 3 kb downstream from the structura l gene produced normal levels of enzyme activity and rescued the poor viabi lity of flies lacking the enzyme. Ail the regulatory elements essential for normal enzyme expression were located in a fragment that included the exon s and introns and 1-kb upstream noncoding sequence. Deletions of the 1.6-kb second intron reduced activity to 25%. Transformants with fusion construct s between the sn-glycerol-3- phosphate dehydrogenase gene and the P-galacto sidase gene from E. coli revealed three elements that affected expression. A (CT)(9) repeat element at the 5' end of the second intron increased expre ssion in both larvae and adults, particularly at emergence. A second regula tory element, which includes a (CT)(7) repeat, was located 5' to the TATA b ox and had similar effects on the gene's expression. A third undefined, enh ancer was located in rite second intron, between 0.5 and 1.8 kb downstream of the translation initiation codon. This element increases enzyme activity to a similar extent in larvae and adults but has little effect when the en hancer at the 5' end of the intron is present.