A highly conserved enhancer in mammalian type X collagen genes drives high levels of tissue-specific expression in hypertrophic cartilage in vitro and in vivo

Sonja Gebhard, Ernst Pöschl, Silvia Riemer, Eva Bauer, Takako Hattori, Heidi Eberspaecher, Zhaoping Zhang, Veronique Lefebvre, Benoit De Crombrugghe, Klaus Von Der Mark

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Previously we have identified a cis-acting regulatory domain in the human type X collagen gene upstream of the transcription start site which acts as a strong enhancer in hypertrophic, but not in resting chondrocytes. Here we show that this enhancer is highly conserved also in the murine and bovine Col10a1 genes, but not found in the known promoter sequences of chicken Col10a1. It contains a functionally active AP-1 site (TPA Responsive Element, TRE) which is essential for the high transcriptional activity of the COL10A1 enhancer in transiently transfected hypertrophic chondrocytes. Gel-shift experiments with nuclear extracts of hypertrophic chondrocytes revealed FosB and Fra-1 as candidates regulating AP-1 factors binding to the TRE site. In fact, coexpression of FosB and Fra-1 in reporter gene assays greatly stimulated transcriptional activity of enhancer bearing reporter genes. Quantitative analysis of AP-1 factor mRNA levels in distinct fractions of fetal bovine epiphyseal chondrocytes by real-time PCR confirmed significant levels of FosB and Fra-1 mRNA besides other AP-1 factors in hypertrophic chondrocytes. A key role of the enhancer element in regulating tissue-specific expression of the Col10a1 gene was shown by establishing transgenic mouse lines with a reporter gene containing a 4.6 kb murine Col10a1 promoter fragment which included the enhancer, exon 1, part of exon 2 and the first intron. Reporter gene expression was seen exclusively in hypertrophic cartilages in the growth plates of long bones, ribs, vertebrae, sternum and mandibles of 17.5-18.5 dpc embryos, confirming that the 4.6 kb promoter is able to drive specific expression of Col10a1 in hypertrophic cartilage. These established transgenic lines should facilitate the genetic analysis of regulatory pathways of chondrocyte maturation and Col10a1 gene expression in the future.

Original languageEnglish
Pages (from-to)309-322
Number of pages14
JournalMatrix Biology
Volume23
Issue number5
DOIs
Publication statusPublished - Aug 2004
Externally publishedYes

Fingerprint

Collagen Type X
Chondrocytes
Cartilage
Transcription Factor AP-1
Reporter Genes
Genes
Gene Expression
Exons
Messenger RNA
Sternum
Growth Plate
Transcription Initiation Site
Ribs
Mandible
Introns
Transgenic Mice
In Vitro Techniques
Real-Time Polymerase Chain Reaction
Chickens
Spine

Keywords

  • Chondrocyte maturation
  • Col10a1 gene expression
  • Exon

ASJC Scopus subject areas

  • Molecular Biology

Cite this

A highly conserved enhancer in mammalian type X collagen genes drives high levels of tissue-specific expression in hypertrophic cartilage in vitro and in vivo. / Gebhard, Sonja; Pöschl, Ernst; Riemer, Silvia; Bauer, Eva; Hattori, Takako; Eberspaecher, Heidi; Zhang, Zhaoping; Lefebvre, Veronique; De Crombrugghe, Benoit; Von Der Mark, Klaus.

In: Matrix Biology, Vol. 23, No. 5, 08.2004, p. 309-322.

Research output: Contribution to journalArticle

Gebhard, S, Pöschl, E, Riemer, S, Bauer, E, Hattori, T, Eberspaecher, H, Zhang, Z, Lefebvre, V, De Crombrugghe, B & Von Der Mark, K 2004, 'A highly conserved enhancer in mammalian type X collagen genes drives high levels of tissue-specific expression in hypertrophic cartilage in vitro and in vivo', Matrix Biology, vol. 23, no. 5, pp. 309-322. https://doi.org/10.1016/j.matbio.2004.05.010
Gebhard, Sonja ; Pöschl, Ernst ; Riemer, Silvia ; Bauer, Eva ; Hattori, Takako ; Eberspaecher, Heidi ; Zhang, Zhaoping ; Lefebvre, Veronique ; De Crombrugghe, Benoit ; Von Der Mark, Klaus. / A highly conserved enhancer in mammalian type X collagen genes drives high levels of tissue-specific expression in hypertrophic cartilage in vitro and in vivo. In: Matrix Biology. 2004 ; Vol. 23, No. 5. pp. 309-322.
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AU - Bauer, Eva

AU - Hattori, Takako

AU - Eberspaecher, Heidi

AU - Zhang, Zhaoping

AU - Lefebvre, Veronique

AU - De Crombrugghe, Benoit

AU - Von Der Mark, Klaus

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