CCAAT binding transcription factor binds and regulates human COL1A1 promoter activity in human dermal fibroblasts: demonstration of increased binding in systemic sclerosis fibroblasts

Arthritis Rheum. 2000 Oct;43(10):2219-29. doi: 10.1002/1529-0131(200010)43:10<2219::AID-ANR9>3.0.CO;2-N.


Objective: To determine the binding factors that interact with the proximal promoter region of the human type I collagen gene, COL1A1, and to examine their involvement in its transcriptional regulation in normal and systemic sclerosis (SSc) dermal fibroblasts.

Methods: Nuclear extracts from dermal fibroblasts from 4 patients with SSc and 4 age- and sex-matched control individuals were examined by electrophoresis mobility shift assays with a COL1A1 promoter fragment encompassing nucleotides -174 to -50 bp. Supershift assays with antibodies specific to various transcription factors, and competition experiments using consensus, wild-type, or mutated oligonucleotides corresponding to their specific binding sites, were performed. The effects of specific oligonucleotides as "intracellular competitors" were examined by transient transfection experiments in SSc fibroblasts using a COL1A1 construct containing -174 bp of the promoter.

Results: The findings demonstrate that the CCAAT binding transcription factor (CBF) binds the proximal CCAAT box located at -100 to -96 bp, but not the distal CCAAT box at -125 to -121 bp, of the human COL1A1 promoter in both SSc and normal fibroblasts. CBF binding activity was 3-5-fold higher in the SSc fibroblasts. Moreover, the promoter activity of the -174-bp COL1A1 construct was decreased by up to 50% when specific oligonucleotides were used as "intracellular competitors." In addition, Sp1 and Sp3 were other transcription factors found to be involved in the formation of the DNA-protein complexes within this region of the COL1A1 promoter.

Conclusion: These results indicate that the transcription factor CBF binds the human COL1A1 proximal promoter region in human dermal fibroblasts, and its binding activity is higher in SSc fibroblasts.

Publication types

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

MeSH terms

  • Base Sequence
  • Binding, Competitive / drug effects
  • CCAAT-Binding Factor / metabolism*
  • CCAAT-Binding Factor / pharmacology*
  • Cells, Cultured
  • Collagen / genetics*
  • DNA-Binding Proteins / pharmacology
  • Fibroblasts / metabolism
  • Gene Expression Regulation / drug effects
  • Humans
  • Oligonucleotide Probes
  • Promoter Regions, Genetic / drug effects
  • Promoter Regions, Genetic / genetics
  • Scleroderma, Systemic / metabolism*
  • Scleroderma, Systemic / pathology*
  • Skin / pathology*
  • Sp1 Transcription Factor* / pharmacology
  • Sp3 Transcription Factor
  • Transcription Factors / metabolism*
  • Transcription Factors / pharmacology


  • CCAAT-Binding Factor
  • DNA-Binding Proteins
  • Oligonucleotide Probes
  • SP3 protein, human
  • Sp1 Transcription Factor
  • Transcription Factors
  • Sp3 Transcription Factor
  • Collagen