Quantitation of glucose-6-phosphate dehydrogenase mRNA by solution hybridization: Correlation with rates of synthesis

Abstract

Rat liver glucose-6-phosphate dehydrogenase (G6PD) is one of several proteins involved in lipid metabolism whose synthesis is regulated by diet. In experiments reported here, rats were fasted or fed diets until a new steady state level of G6PD was produced. Livers were used to measure G6PD activity, synthesis and mRNA simultaneously. Since accurate quantitation of G6PD mRNA by Northern blots was found to be difficult in noninduced animals a new solution hybridization assay was also used. Noninduced rats have approx. One molecule of G6PD mRNA per liver cell. Changes in G6PD mRNA are larger than previously reported and, at the steady state, can completely account for the 33-fold change in G6PD activity and synthesis when fasted rats are refed a high carbohydrate diet. In contrast, a high fat carbohydrate-free diet does not increase G6PD mRNA and dibutyryl cAMP lowers G6PD mRNA. Since changes in G6PD synthesis and activity are closely correlated, degradation of G6PD is not significantly regulated.