5’ AMPK Gene Family: Exploration of Conserved DNA Methylation

The AMPK gene family is responsible for cellular metabolism and energy regulation. Activation of these genes is caused by stress induced environmental and nutritional factors. The primary function of these genes is to provide energy-conserving measures when ATP levels are being depleted. Epigenetic modifications of DNA methylation have been associated with the regulation of this gene family. DNA methylation involves the addition of a methyl group onto the 5 carbon position of cytosines within CpG dinucleotides. The presence of methylation in the promoter region of a gene can prevent transcription and thus alter phenotypic effects. Of those seven genes that comprise the AMPK gene family, five genes were selected to undergo Combined Bisulfite Restriction Analysis (COBRA). Primers pairs for each of these five genes were generated with MethPrimer and used to amplify DNA extracted from the liver and muscle tissues of two breeds of cattle, Angus (n=6) and Charolais (n=6). The presence of DNA methylation was initially determined by combined bisulfite restriction analysis and positive results were confirmed with Sanger sequencing. Sequencing results showed conservation of DNA methylation in the bovine AMPK gene family and across bovine breeds and tissues. In order to determine the evolutionary pattern of conserved DNA methylation, our next step is to expand our samples to include additional breeds and species, including Bos indicus, Bison bison and Alces alces.