Analysis of regulatory regions in the COL1A1 gene responsible for 1,25-Dihydroxyvitamin D3-mediated transcriptional repression in osteoblastic cells

D. Pavlin, A. Bedalov, M. S. Kronenberg, B. E. Kream, D. W. Rowe, Catharine Smith, J. W. Pike, A. C. Lichtler

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The synthesis of type I collagen in bone cells is inhibited by the calcium-regulating hormone 1,25-dihydroxyvitamin D3. Earlier work from our laboratories has indicated that vitamin D regulation is at the level of transcription, based on results from both nuclear run-off assays and functional promoter analysis of a hybrid gene consisting of a 3.6 kb COL1A1 promoter fragment fused to the chloramphenicol acetyltransferase reporter gene. In the present study, we investigated the molecular basis for vitamin D-mediated transcriptional repression of the COL1A1 gene and report the identification of a region within the COL1A1 upstream promoter (the HindIII-PstI restriction fragment between nucleotides -2295 and -1670) which is necessary for 1,25-dihydroxyvitamin D3 responsiveness in osteoblastic cells. This hormone-mediated inhibitory effect on the marker gene parallels the inhibition of the endogenous collagen gene. A 41 bp fragment from this region (between nucleotides -2256 and -2216) contains a sequence which is very similar to vitamin D-responsive elements identified in the osteocalcin gene. Extracts from cultured cells which express a high level of vitamin D receptor contain a hormone:receptor complex that binds specifically to this 41 bp fragment, as demonstrated by bandshift analysis. However, deletion of this vitamin D receptor binding region from either a -3.5 kb or a -2.3 kb promoter fragment did not abolish vitamin D responsiveness, These results indicate that a vitamin D response element similar to that described for other vitamin D responsive genes (osteocalcin and osteopontin) does not alone mediate the repression of COL1A1 by 1,25-dihydroxyvitamin D3.

Original languageEnglish (US)
Pages (from-to)490-501
Number of pages12
JournalJournal of Cellular Biochemistry
Volume56
Issue number4
DOIs
StatePublished - 1994
Externally publishedYes

Fingerprint

Calcitriol
Nucleic Acid Regulatory Sequences
Vitamin D
Genes
Calcitriol Receptors
Osteocalcin
Hormones
Vitamin D Response Element
Nucleotides
Osteopontin
Chloramphenicol O-Acetyltransferase
Functional analysis
Collagen Type I
Reporter Genes
Transcription
Cultured Cells
Collagen
Assays
Bone
Calcium

Keywords

  • Bone cells in culture
  • Gene regulation by steroid hormone
  • Nucleotides
  • Type I collagen
  • Vitamin D

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Analysis of regulatory regions in the COL1A1 gene responsible for 1,25-Dihydroxyvitamin D3-mediated transcriptional repression in osteoblastic cells. / Pavlin, D.; Bedalov, A.; Kronenberg, M. S.; Kream, B. E.; Rowe, D. W.; Smith, Catharine; Pike, J. W.; Lichtler, A. C.

In: Journal of Cellular Biochemistry, Vol. 56, No. 4, 1994, p. 490-501.

Research output: Contribution to journalArticle

Pavlin, D. ; Bedalov, A. ; Kronenberg, M. S. ; Kream, B. E. ; Rowe, D. W. ; Smith, Catharine ; Pike, J. W. ; Lichtler, A. C. / Analysis of regulatory regions in the COL1A1 gene responsible for 1,25-Dihydroxyvitamin D3-mediated transcriptional repression in osteoblastic cells. In: Journal of Cellular Biochemistry. 1994 ; Vol. 56, No. 4. pp. 490-501.
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abstract = "The synthesis of type I collagen in bone cells is inhibited by the calcium-regulating hormone 1,25-dihydroxyvitamin D3. Earlier work from our laboratories has indicated that vitamin D regulation is at the level of transcription, based on results from both nuclear run-off assays and functional promoter analysis of a hybrid gene consisting of a 3.6 kb COL1A1 promoter fragment fused to the chloramphenicol acetyltransferase reporter gene. In the present study, we investigated the molecular basis for vitamin D-mediated transcriptional repression of the COL1A1 gene and report the identification of a region within the COL1A1 upstream promoter (the HindIII-PstI restriction fragment between nucleotides -2295 and -1670) which is necessary for 1,25-dihydroxyvitamin D3 responsiveness in osteoblastic cells. This hormone-mediated inhibitory effect on the marker gene parallels the inhibition of the endogenous collagen gene. A 41 bp fragment from this region (between nucleotides -2256 and -2216) contains a sequence which is very similar to vitamin D-responsive elements identified in the osteocalcin gene. Extracts from cultured cells which express a high level of vitamin D receptor contain a hormone:receptor complex that binds specifically to this 41 bp fragment, as demonstrated by bandshift analysis. However, deletion of this vitamin D receptor binding region from either a -3.5 kb or a -2.3 kb promoter fragment did not abolish vitamin D responsiveness, These results indicate that a vitamin D response element similar to that described for other vitamin D responsive genes (osteocalcin and osteopontin) does not alone mediate the repression of COL1A1 by 1,25-dihydroxyvitamin D3.",
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AU - Kream, B. E.

AU - Rowe, D. W.

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AU - Pike, J. W.

AU - Lichtler, A. C.

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