Identification of transcription factors involved in the transcriptional regulation of the CXCL12 gene in rat pancreatic insulinoma Rin-5F cell line

Biochem Cell Biol. 2015 Feb;93(1):54-62. doi: 10.1139/bcb-2014-0104. Epub 2014 Sep 29.

Abstract

Diabetes is characterized by a deficit in the number of functional pancreatic β-cells. Understanding the mechanisms that stimulate neogenesis of β-cells should contribute to improved maintenance of β-cell mass. Chemokine CXCL12 has recently become established as a novel β-cell growth factor, however the mechanisms controlling its expression require clarification. We investigated the proteins involved in the transcriptional regulation of the rat β-cell CXCL12 gene (Cxcl12). Using the electrophoretic mobility shift assay and chromatin immunoprecipitation, we established the in vitro and in vivo binding of C/EBPβ, C/EBPα, STAT3, p53, FOXO3a, and HMG I/Y to the Cxcl12 promoter. Co-immunoprecipitation experiments revealed protein-protein interactions between YY1 and PARP-1, FOXO3a and PARP-1, Sp1 and PARP-1, p53 and PARP-1, C/EBPβ and PARP-1, YY1 and p53, YY1 and FOXO3a, p53 and FOXO3a, Sp1 and FOXO3a, C/EBPβ and FOXO3a, C/EBPα and FOXO3a, Sp1 and STAT3. Our data lay the foundation for research into the interplay of signaling pathways that determine the β-cell Cxcl12 expression profile.

Keywords: CXCL12; CXCL12 gene promoter; interaction facteur de transcription-promoteur; interaction protéine–protéine; promoteur du gène de CXCL12; protein–protein interaction; régulation transcriptionnelle; transcription factor-promoter interaction; transcriptional regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • CCAAT-Enhancer-Binding Protein-beta / genetics
  • CCAAT-Enhancer-Binding Protein-beta / metabolism*
  • Cell Line, Tumor
  • Chemokine CXCL12 / genetics*
  • Chromatin Immunoprecipitation
  • Electrophoretic Mobility Shift Assay
  • Gene Expression Regulation
  • Insulin-Secreting Cells / metabolism*
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Promoter Regions, Genetic
  • Rats
  • STAT3 Transcription Factor / genetics
  • STAT3 Transcription Factor / metabolism*
  • Transcriptional Activation

Substances

  • CCAAT-Enhancer-Binding Protein-beta
  • CXCL12 protein, rat
  • Cebpb protein, rat
  • Chemokine CXCL12
  • STAT3 Transcription Factor
  • Stat3 protein, rat
  • Poly(ADP-ribose) Polymerases