The role of methylation of glutamate dehydrogenase gene promoters (<i>GDH1</i> and <i>GDH2</i>) in the regulation of their expression in corn leaves under hypoxia

Abstract

The regulation of glutamate dehydrogenase, an enzyme that is involved in both nitrogen and carbon metabolism, and also links between the tricarboxylic acid cycle and the -aminobutyric acid shunt, has been studied. It was found that oxygen deficiency-induced changes in glutamate dehydrogenase activity in maize leaves (Zea mays L.) are to increase its catalytic activity by more than twice. Differential expression of genes was studied by real-time PCR in GDH1 and GDH2, which encode the - and -subunits of glutamate dehydrogenase, respectively, in the maize genome. Decreased relative level of gene transcripts GDH2 was accompanied by an increase in the expression activity of the gene GDH1. This, in turn, presumably promoted the amination reaction of 2-oxoglutarate. In the promoter of the gene GDH2, the presence of two CpG islands 404 and 383 bp in size was found. Gene promoter GDH1 does not contain a single CpG island; however, 38% of the CpNpG and CpNpN sites of the total number of studied dinucleotides in its composition were found. To assess the influence of the degree of methylation of individual CpG dinucleotides that are part of the promoter regions of genes GDH1 and GDH2 on their expression under hypoxic conditions, a comparative analysis of the dynamics of the transcriptional activity of the genes of - and -subunits of glutamate dehydrogenase from the methyl status of their promoters was carried out. Inversely proportional superposition of changes in the methylation profile of gene promoters GDH1 and GDH2 and transformation of the level of expression of these genes shows their correlation. The data obtained as a result of methyl-specific PCR indicate that an increase in the proportion of methylated CpG dinucleotides leads to a decrease in the amount of mRNA of the gene GDH2, while a decrease in this value for the gene GDH1 causes the induction of its functioning. Methylation of promoter regions of glutamate dehydrogenase genes regulates their transcriptional activity in maize leaves in vivo under conditions of oxygen deficiency. Thus, the little data on the molecular mechanisms of regulation of the synthesis of glutamate dehydrogenase isoenzymes were supplemented by new results on the role of the degree of methylation of gene promoters GDH1 and GDH2 glutamate dehydrogenases in their differential expression during maizes adaptation to hypoxia.