TY - JOUR
T1 - DNA binding of the p21 repressor ZBTB2 is inhibited by cytosine hydroxymethylation
AU - Lafaye, Céline
AU - Barbier, Ewa
AU - Miscioscia, Audrey
AU - Saint-Pierre, Christine
AU - Kraut, Alexandra
AU - Couté, Yohann
AU - Plo, Isabelle
AU - Gasparutto, Didier
AU - Ravanat, Jean Luc
AU - Breton, Jean
N1 - Funding Information:
This work was supported by the Agence Nationale de la Recherche (Epigenome project, coordinator: Murat Saparbaev, Institut Gustave Roussy). We thank the Membrane Protein Purification Platform, from the Institut de Biologie Structurale in Grenoble, for the purification of MeCP2. We thank L. Matthys and C. Biros for the verification of English spelling.
PY - 2014/3/28
Y1 - 2014/3/28
N2 - 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.
AB - 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.
KW - 5-hydroxymethylcytosine
KW - Protein-DNA interactions
KW - ZBTB2
UR - http://www.scopus.com/inward/record.url?scp=84897986932&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2014.02.122
DO - 10.1016/j.bbrc.2014.02.122
M3 - Article
C2 - 24607898
AN - SCOPUS:84897986932
SN - 0006-291X
VL - 446
SP - 341
EP - 346
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -