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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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
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author Szwagierczak, Aleksandra
Brachmann, Andreas
Schmidt, Christine S.
Bultmann, Sebastian
Leonhardt, Heinrich
Spada, Fabio
author_facet Szwagierczak, Aleksandra
Brachmann, Andreas
Schmidt, Christine S.
Bultmann, Sebastian
Leonhardt, Heinrich
Spada, Fabio
author_sort Szwagierczak, Aleksandra
collection PubMed
description 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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spelling pubmed-31302832011-07-06 Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine Szwagierczak, Aleksandra Brachmann, Andreas Schmidt, Christine S. Bultmann, Sebastian Leonhardt, Heinrich Spada, Fabio Nucleic Acids Res Nucleic Acid Enzymes 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. Oxford University Press 2011-07 2011-03-04 /pmc/articles/PMC3130283/ /pubmed/21378122 http://dx.doi.org/10.1093/nar/gkr118 Text en © The Author(s) 2011. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Nucleic Acid Enzymes
Szwagierczak, Aleksandra
Brachmann, Andreas
Schmidt, Christine S.
Bultmann, Sebastian
Leonhardt, Heinrich
Spada, Fabio
Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine
title Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine
title_full Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine
title_fullStr Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine
title_full_unstemmed Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine
title_short Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine
title_sort characterization of pvurts1i endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine
topic Nucleic Acid Enzymes
url 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
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