Exploring DNA Methylation in the Bovine 5’ AMPK Gene Family

The 5’ AMP-activated protein kinase AMPK gene family plays a primary function in regulating cellular energy and metabolism. The activation of this gene family primarily occurs when there is a depletion of cellular ATP due to environmental and nutritional stressors, which is associated with a rise of AMP levels.  Subsequently, the genes initiate energy-conserving measures within the cell to protect it from the decrease in ATP levels.  The regulation of these genes has been linked to the epigenetic mechanism of methylation.   DNA methylation primarily involves the addition of a methyl group to the 5 position of cytosines that are found in CpG dinucleotides.  When 5-methylcytosine is found in the 5’ promoter region of a gene, it can prevent transcriptional machinery from binding to its target site resulting in the silencing of gene transcription.  We have investigated the presence of methylation in five of the seven genes that comprise the AMPK gene family.  From these 5 genes, a total of 13 primer pairs were designed using MethPrimer, in which 11 primer pairs could be optimized.  These primer pairs have been used to amplify the DNA extracted from liver and muscle tissue in each of six Angus and Charolais cattle (3 High RFI, 3 Low RFI). Levels of DNA methylation were determined by amplification of bisulfite treated DNA followed by combined, bisulfite restriction analysis (COBRA).  This study confirms the presence of DNA methylation in the bovine AMPK gene family and provides evidence of conservation of methylation across breeds of cattle.