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Papers In Press, published online ahead of print June 18, 2002
Biochemistry and Molecular Pharmacology, West Virginia University, Morgantown, WV 26506-9142
Corresponding Author: LSALATI{at}hsc.wvu.edu
Polyunsaturated fatty acids inhibit the expression of hepatic glucose-6-phosphate dehydrogenase (G6PD) by changes in the amount of G6PD pre-mRNA in the nucleus in the absence of changes in the transcription rate of the gene. We have compared the nuclear accumulation of partially and fully spliced mRNA for G6PD in livers of mice fed diets high versus low in polyunsaturated fat. Consumption of a diet high in polyunsaturated fat decreased the accumulation of partially spliced forms of the G6PD pre-mRNA. Examining the fate of multiple introns within the G6PD primary transcript indicated that in mice fed a high fat diet, G6PD pre-mRNA containing intron 11 accumulated within the nucleus while G6PD mature mRNA abundance was inhibited 50% or more within the same livers. Transient transfection of RNA reporters into primary hepatocyte cultures was used to localize the cis-acting RNA element involved in this regulated splicing. Reporter RNA produced from constructs containing exon 12 were decreased in amount by arachidonic acid. The extent of this decrease paralleled that seen in the expression of the endogenous G6PD mRNA. The presence of both exon 12 and a neighboring intron within the G6PD reporter RNA was essential for regulation by polyunsaturated fatty acid. Inhibition was not dependent on the presence of the G6PD polyadenylation signal and 3’-UTR, but substitution with the SV40 poly (A) signal attenuated the inhibition by arachidonic acid. Thus exon 12 contains a putative splicing regulatory element involved in the inhibition of G6PD expression by polyunsaturated fat.
J. Biol. Chem, 10.1074/jbc.M203196200
Submitted on April 3, 2002
Revised on June 18, 2002
Accepted on June 18, 2002
Inhibition of the splicing of glucose-6-phosphate dehydrogenase pre-mRNA by polyunsaturated fatty acids
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