For more than a decade, DNA and histone methylations have been the focus of extensive work, although their relationship with methyl group metabolism was overlooked. Recently, it has emerged that epigenetic methylations are influenced by methyl donor nutrient availability, cellular levels of S-adenosyl-methionine (SAM), and cytoplasmic methyltransferase activities. SAM-dependent methyltransferases methylate a wide range of targets, from small molecules to proteins and nucleic acids. However, few investigations of the global methylome of tumors have been performed. Here, untargeted NMR metabolomics of two mouse tumor models labeled with [13C-methyl]methionine were used to search for the NMR-visible set of cellular methyl acceptors denoted the global methylome. Tumor models were B16 melanoma cell cultures and B16 melanoma tumors, which may be considered as two stages of B16 tumor development. Based on 2D 1H-13C NMR spectra and orthogonal partial least squares discriminant analysis of spectra, our study revealed markedly different global methylomes for melanoma models. The methylome of B16 melanoma cell cultures was dominated by histone methylations, whereas that of B16 melanoma tumors was dominated by cytoplasmic small-molecule methylations. Overall, the technique gave access to the non-DNA methylome. Comparison of tumor models also exhibiting differential expression of aerobic glycolysis provided clues to a methyl metabolism shift during tumor progression.
Keywords: 2D 1H−13C NMR spectroscopy; B16 melanoma cell culture; B16 melanoma tumor; global methylome; high-resolution magic angle spinning; l-[13C-methyl]methionine; metabolomics; methyl metabolism; orthogonal partial least squares discriminant analysis.