GATA-4 binds to an upstream element of the human α2(I) collagen gene (COL1A2) and inhibits transcription in fibroblasts

  • Wang L
  • Tanaka S
  • Ramirez F
  • 3


    Mendeley users who have this article in their library.
  • 13


    Citations of this article.


Chromatin analyses have identified two DNase I hypersensitive sites (HS1 and HS2) at comparable distances (- 130 bp and -2.3 kb) from the transcription start site of the human and mouse α2(I) collagen gene. Whereas the DNA region encompassing HS1 has been extensively characterized using protein binding and functional assays, nothing is yet known about the contribution of the HS2 sequence to α2(I) collagen gene transcription. Here we report that the HS2 sequence of the human α2(I) collagen (COL1A2) gene is a binding site for a transcriptional repressor in fibroblasts. DNase I footprinting identified a single site of nuclease protection around HS2, which corresponds to a sequence potentially capable of forming a 13 bp long hairpin structure with a 4 bp interruption in the middle. Gel mobility shift assays revealed that two GATA consensus sequences embedded within the hairpin are specifically bound by GATA-4 in fibroblasts. They also showed that formation of the HS2 protein complex requires the integrity of the whole hairpin sequence. Transient transfections of luciferase reporter gene constructs in fibroblasts correlated the HS2 element with transcriptional repression of the - 2.3 kb promoter sequence. This last observation was further corroborated by showing that forced overexpression of GATA-4 in cultured fibroblasts leads to decreased transcription from the co-transfected - 2.3 kb promoter/reporter construct, as well as reduced expression of the endogenous collagen gene. Finally, a chromatin immunoprecipitation assay documented GATA-4 ability to bind to the HS2 element in vivo. These results are therefore the first to implicate GATA-4 in regulating constitutive COL1A2 gene expression in fibroblasts. © 2005 Elsevier B.V./International Society of Matrix Biology. All rights reserved.

Author-supplied keywords

  • COL1A2 transcription
  • Collagen regulation
  • GATA
  • Transcriptional repression

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document


  • Lu Wang

  • Shizuko Tanaka

  • Francesco Ramirez

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free