The effect of DNA damage on the pattern of immune-detectable DNA methylation in mouse embryonic fibroblasts

摘要: The methylation of cytosine at CpG dinucleotides (5 meC) is an important epigenetic mechanism that governs genome stability and gene expression. Important ontological pathological transitions are associated with marked global changes in detectable levels methylation. We have previously found two pools immune-detectable 5 meC exist within cells, a pool can be detected after acid treatment fixed cells to denature chromatin another large but variable requires further tryptic digestion step for complete epitope retrieval. trypsin-sensitive has been shown largely the heterochromatic fraction (by heterochromatin marker, HP1-β) genome, size this varies growth disposition cells. Since DNA damage imposes on structure present study analyzed how such influences faithful immunological detection mouse embryonic fibroblasts. was induced by either UV-irradiation or doxorubicin treatment, each which resulted increased 24-48 h treatment. There these significantly damage. staining predominantly co-located foci nuclei, as assessed HP1-β staining. relative amount masked positively HP1-β. methyl binding protein, MBD1, less reliable measure meC, significant not being MBD1. cyto-epigenetic approaches used here reveal dynamism localization nuclei fibroblasts response