Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine

In mammalian genomes a sixth base, 5-hydroxymethylcytosine ((hm)C), is generated by enzymatic oxidation of 5-methylcytosine ((m)C). This discovery has raised fundamental questions about the functional relevance of (hm)C in mammalian genomes. Due to their very similar chemical structure, discriminati...

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Detalles Bibliográficos
Autores principales: Szwagierczak, Aleksandra, Brachmann, Andreas, Schmidt, Christine S., Bultmann, Sebastian, Leonhardt, Heinrich, Spada, Fabio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2011
Materias:
Nucleic Acid Enzymes
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3130283/
https://www.ncbi.nlm.nih.gov/pubmed/21378122
http://dx.doi.org/10.1093/nar/gkr118
Descripción
Sumario:In mammalian genomes a sixth base, 5-hydroxymethylcytosine ((hm)C), is generated by enzymatic oxidation of 5-methylcytosine ((m)C). This discovery has raised fundamental questions about the functional relevance of (hm)C in mammalian genomes. Due to their very similar chemical structure, discrimination of the rare (hm)C against the far more abundant (m)C is technically challenging and to date no methods for direct sequencing of (hm)C have been reported. Here, we report on a purified recombinant endonuclease, PvuRts1I, which selectively cleaves (hm)C-containing sequences. We determined the consensus cleavage site of PvuRts1I as (hm)CN(11–12)/N(9–10)G and show first data on its potential to interrogate (hm)C patterns in mammalian genomes.

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