A novel cis-element that enhances connective tissue growth factor gene expression in chondrocytic cells

Biochem Biophys Res Commun. 2002 Jul 12;295(2):445-51. doi: 10.1016/s0006-291x(02)00700-3.

Abstract

To clarify the chondrocyte-specific regulatory mechanism of connective tissue growth factor (ctgf) gene expression, we analyzed the functionality and DNA-protein interaction of the CTGF promoter. Comparative luciferase assay of the CTGF promoter deletion mutants among HCS-2/8 chondrocytic cells and fibroblastic cells revealed that a 110-bp region in the promoter was crucial for the HCS-2/8-specific transcriptional enhancement. Subsequent competitive gel shift assay revealed that transcription factors in HCS-2/8 nuclei bound to a 60-bp portion in the corresponding region. Relative luciferase activity from a CTGF promoter with mutant TGF-beta response element (TbRE) was 16.9% lower than that from an intact promoter. On the other hand, relative luciferase activity from a CTGF promoter with 4bp point mutations at 30bp upstream of the TbRE was 47.7% lower than that from the intact one. The binding activity of HCS-2/8 nuclear factor(s) to the sequence over the 4-bp was remarkably higher than that of any nuclear extract from other types of cells. Therefore, we entitled the sequence 'TRENDIC', a transcription enhancer dominant in chondrocytes, which stands for a novel enhancer for chondrocyte-specific CTGF gene expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Cell Line
  • Chondrocytes / metabolism*
  • Connective Tissue Growth Factor
  • DNA
  • Enhancer Elements, Genetic
  • Gene Expression Regulation / physiology*
  • Growth Substances / genetics*
  • Growth Substances / metabolism
  • Humans
  • Immediate-Early Proteins / genetics*
  • Immediate-Early Proteins / metabolism
  • Intercellular Signaling Peptides and Proteins*
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Transcription Factors / metabolism

Substances

  • CCN2 protein, human
  • Growth Substances
  • Immediate-Early Proteins
  • Intercellular Signaling Peptides and Proteins
  • Transcription Factors
  • Connective Tissue Growth Factor
  • DNA