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

Szwagierczak, Aleksandra; Brachmann, Andreas ; Schmidt, Christine S.; Bultmann, Sebastian; Leonhardt, Heinrich und Spada, Fabio (1. Juli 2011): Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5–hydroxymethylcytosine. In: Nucleic Acids Research, Bd. 39, Nr. 12: S. 5149-5156

Abstract

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.

• Dokumententyp: Zeitschriftenartikel
• Publikationsform: Publisher's Version
• Fakultät: Biologie > Department Biologie I
• Themengebiete: 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie
• ISSN: 0305-1048
• Sprache: Englisch