DNA binding of the p21 repressor ZBTB2 is inhibited by cytosine hydroxymethylation.

Céline Lafaye, Ewa Barbier, Audrey Miscioscia, Christine Saint-Pierre, Alexandra Kraut, Yohann Couté, Isabelle Plo, Didier Gasparutto, Jean-Luc Ravanat, Jean Breton

Biochemical and biophysical research communications 2014 Mar 28

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Recent studies have demonstrated that the modified base 5-hydroxymethylcytosine (5-hmC) is detectable at various rates in DNA extracted from human tissues. This oxidative product of 5-methylcytosine (5-mC) constitutes a new and important actor of epigenetic mechanisms. We designed a DNA pull down assay to trap and identify nuclear proteins bound to 5-hmC and/or 5-mC. We applied this strategy to three cancerous cell lines (HeLa, SH-SY5Y and UT7-MPL) in which we also measured 5-mC and 5-hmC levels by HPLC-MS/MS. We found that the putative oncoprotein Zinc finger and BTB domain-containing protein 2 (ZBTB2) is associated with methylated DNA sequences and that this interaction is inhibited by the presence of 5-hmC replacing 5-mC. As published data mention ZBTB2 recognition of p21 regulating sequences, we verified that this sequence specific binding was also alleviated by 5-hmC. ZBTB2 being considered as a multifunctional cell proliferation activator, notably through p21 repression, this work points out new epigenetic processes potentially involved in carcinogenesis. Copyright © 2014 Elsevier Inc. All rights reserved.

Citation

Céline Lafaye, Ewa Barbier, Audrey Miscioscia, Christine Saint-Pierre, Alexandra Kraut, Yohann Couté, Isabelle Plo, Didier Gasparutto, Jean-Luc Ravanat, Jean Breton. DNA binding of the p21 repressor ZBTB2 is inhibited by cytosine hydroxymethylation. Biochemical and biophysical research communications. 2014 Mar 28;446(1):341-6