doiSerbia

Enrichment of Cxcl12 promoter with TET2: A possible link between promoter demethylation and enhanced gene expression in the absence of PARP-1

Tolić Anja Z. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Rajić Jovana J. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Đorđević Marija B. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Đorđević Miloš M. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Dinić Svetlana S. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Grdović Nevena M. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Arambašić-Jovanović Jelena D. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Mihailović Mirjana V. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Poznanović Goran Đ. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Jurkowski Tomasz P. (Institute of Biochemistry, University of Stuttgart, Stuttgart, Germany + School of Biosciences, Cardiff University, Cardiff, Wales, Sir Martin Evans Building, Cardiff, CF AX, UK)
Vidaković Melita S. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)
Uskoković Aleksandra S. (Institute for Biological Research “Siniša Stanković“, Department of Molecular Biology, Belgrade)

Previously, we described the link between C-X-C motif chemokine 12 (Cxcl12) gene induction and DNA hypomethylation in the absence of poly(ADP-ribose) polymerase 1 (PARP-1). We have now firmly established that demethylation is the primary cause of gene induction on the basis of Cxcl12 gene upregulation upon treatment with the demethylating agent 5-azacytidine (5-aza). Since the demethylation state of Cxcl12 is favored by PARP-1 absence, we investigated the presence of ten-eleven translocation (TET) proteins on the Cxcl12 promoter in order to corroborate the relationship between the demethylation process and increased gene expression that occurs in the absence of PARP-1. Analysis was performed on the promoter region within CpG islands of Cxcl12 from control mouse embryonic fibroblasts (NIH3T3) and PARP-1 knock-out mouse embryonic fibroblasts (PARP1-/-). The lack of PARP-1 increased the abundance of TET2 on the Cxcl12 promoter, suggesting that TET-mediated demethylation provoked by the absence of PARP-1 could account for the observed increased expression of this chemokine. Deciphering the regulation of DNA (de)methylation factors that control Cxcl12 expression may provide an additional therapeutic approach in pharmacological interventions where gene switching on or off based on targeted stimulation or inhibition is necessary.

Keywords: DNA demethylation, 5-aza, TET2, PARP-1, CXCL12

Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173020