A novel transforming growth factor beta response element controls the expression of the connective tissue growth factor gene

Cell Growth Differ. 1996 Apr;7(4):469-80.


We reported previously that transforming growth factor beta (TGF-beta) selectively induced high levels of connective tissue growth factor (CTGF) mRNA and protein in human skin fibroblasts. In this study, we investigated the molecular mechanism for TGF-beta regulation of CTGF gene expression. Northern blot and run-on transcription assays indicate that TGF-beta directly activates transcription of the CTGF gene. Fragments of the 5'flanking region of the human CTGF gene were linked to luciferase reporter constructs. TGF-beta induced a 25-30 fold increase in luciferase activity in NIH/3T3 fibroblasts that had been transfected with this construct compared with nontreated cells after 24 h incubation. Other growth factors, such as platelet derived growth factor or fibroblast growth factor, caused only a 2-3-fold induction. This response to TGF-beta occurred only in human skin fibroblasts, fetal bovine aortic smooth muscle cells, and NIH/3T3 fibroblasts but not in the epithelial cell lines tested. Analysis of deletion mutants indicated that an important TGF-beta regulatory element is located between positions -162 and -128 of the CTGF promoter sequence. A fragment of the promoter containing this region conferred TGF-beta induction to a SV40 enhanceriess promoter. Methylation interference and competition gel shift assays mapped a unique 13-nucleotide sequence delineating a novel TGF-beta cis-regulatory element. Point mutations in this region result in a complete loss of the TGF-beta induction, identifying this sequence as a new TGF-beta response element.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Animals
  • Anisomycin / pharmacology
  • Base Sequence
  • Binding Sites
  • Blotting, Northern
  • Connective Tissue Growth Factor
  • Cycloheximide / pharmacology
  • Fibroblasts
  • Gene Deletion
  • Gene Expression Regulation*
  • Growth Substances / genetics*
  • Growth Substances / metabolism
  • Humans
  • Immediate-Early Proteins*
  • Intercellular Signaling Peptides and Proteins*
  • Luciferases
  • Mice
  • Mitogens / genetics*
  • Molecular Sequence Data
  • Oligonucleotides / pharmacology
  • Promoter Regions, Genetic / drug effects
  • Promoter Regions, Genetic / genetics
  • Protein Binding
  • Protein Synthesis Inhibitors / pharmacology
  • Puromycin / pharmacology
  • RNA, Messenger / metabolism
  • Transcription, Genetic
  • Transfection
  • Transforming Growth Factor beta / genetics
  • Transforming Growth Factor beta / pharmacology*


  • CCN2 protein, human
  • CCN2 protein, mouse
  • Growth Substances
  • Immediate-Early Proteins
  • Intercellular Signaling Peptides and Proteins
  • Mitogens
  • Oligonucleotides
  • Protein Synthesis Inhibitors
  • RNA, Messenger
  • Transforming Growth Factor beta
  • Connective Tissue Growth Factor
  • Puromycin
  • Anisomycin
  • Cycloheximide
  • Luciferases