Ultrastructural study of the main epigenetic markers in hepatocyte nuclei
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DNA methylation is one of the most frequently investigated epigenetic processes and is involved in many biological mechanisms. It consists in the addition of a methyl group, donated by the S-adenosyl methionine, on the cytosine carbon 5 to form 5-methylcytosine (5mC). Cytosine methylation is reversible, as 5mC may be converted into 5-hydroxymethylcytosine (5hmC) through an oxidation step. The presence of 5mC and 5hmC has been investigate also on RNA molecules and they have been found in tRNAs, rRNAs and in pre-mature and mature mRNAs. In this work, we studied the distribution of 5mC and 5hmc on DNA and RNA in mouse hepatocyte nuclei by using morphological and morphometric analyses at transmission electron microscopy and a semi-quantitative analysis on extracted mRNA. 5mC and 5hmC were found to occur in different amounts in heterochromatin and perichromatin region (i.e., where euchromatin is located); moreover, their relative amounts differed in DNA and RNA. Our results support the hypothesis that the process of methylation plays a crucial role in gene silencing and expression. Methylation and de-methylation might also represent two preliminary steps in the processing of mRNA, maybe marking the mRNA fraction destined to degradation or that would never leave the nucleus for translation.
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