1,25-Dihydroxyvitamin D3 inhibition of Col1a1 promoter expression in calvariae from neonatal transgenic mice

Antonio Bedalov, Roberto Salvatori, Milan Dodig, Belinda Kapural, Dubravko Pavlin, Barbara E. Kream, Stephen H. Clark, Charles O. Woody, David W. Rowe, Alexander C. Lichtler

Research output: Contribution to journalArticle

Abstract

We studied the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on organ cultures of transgenic mouse calvariae containing segments of the Col1a1 promoter extending to -3518, -2297, -1997, -1794, -1763, and -1719 bp upstream of the transcription start site fused to the chloramphenicol acetyltransferase (CAT) reporter gene. 1,25(OH)2D3 had a dose-dependent inhibitory effect on the expression of the -3518 bp promoter construct (ColCAT3.6), with maximal inhibition of about 50% at 10 nM. This level of inhibition was consistent with the previously observed effect on the endogenous Col1a1 gene in bone cell models. All of the shorter constructs were also inhibited by 10 nM 1,25(OH)2D3, suggesting that the sequences required for 1,25(OH)2D3 inhibition are downstream of -1719 bp. The inhibitory effect of 1,25(OH)2D3 on transgene mRNA was maintained in the presence of the protein synthesis inhibitor cycloheximide, suggesting that the inhibitory effect on Col1a1 gene transcription does not require de novo protein synthesis. We also examined the in vivo effect of 1,25(OH)2D3 treatment of transgenic mice on ColCAT activity, and found that 48 h treatment caused a dose-dependent inhibition of CAT activity in calvariae comparable to that observed in organ cultures. In conclusion, we demonstrated that 1,25(OH)2D3 inhibits Col1A1 promoter activity in transgenic mouse calvariae, both in vivo and in vitro. The results indicate that there is a 1,25(OH)2D3 responsive element downstream of -1719 bp. The inhibitory effect does not require new protein synthesis.

Original languageEnglish (US)
Pages (from-to)285-293
Number of pages9
JournalBBA - Gene Structure and Expression
Volume1398
Issue number3
DOIs
StatePublished - Jul 9 1998

Fingerprint

Calcitriol
Skull
Transgenic Mice
Chloramphenicol O-Acetyltransferase
Genes
Organ Culture Techniques
Protein Synthesis Inhibitors
Transcription Initiation Site
Transcription
Cycloheximide
Transgenes
Reporter Genes
Bone
Proteins
Bone and Bones
Messenger RNA

Keywords

  • Bone
  • Calcium regulating hormone
  • Collagen
  • Osteoblast
  • Transcription

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Structural Biology
  • Biophysics

Cite this

1,25-Dihydroxyvitamin D3 inhibition of Col1a1 promoter expression in calvariae from neonatal transgenic mice. / Bedalov, Antonio; Salvatori, Roberto; Dodig, Milan; Kapural, Belinda; Pavlin, Dubravko; Kream, Barbara E.; Clark, Stephen H.; Woody, Charles O.; Rowe, David W.; Lichtler, Alexander C.

In: BBA - Gene Structure and Expression, Vol. 1398, No. 3, 09.07.1998, p. 285-293.

Research output: Contribution to journalArticle

Bedalov, A, Salvatori, R, Dodig, M, Kapural, B, Pavlin, D, Kream, BE, Clark, SH, Woody, CO, Rowe, DW & Lichtler, AC 1998, '1,25-Dihydroxyvitamin D3 inhibition of Col1a1 promoter expression in calvariae from neonatal transgenic mice', BBA - Gene Structure and Expression, vol. 1398, no. 3, pp. 285-293. https://doi.org/10.1016/S0167-4781(98)00079-7
Bedalov, Antonio ; Salvatori, Roberto ; Dodig, Milan ; Kapural, Belinda ; Pavlin, Dubravko ; Kream, Barbara E. ; Clark, Stephen H. ; Woody, Charles O. ; Rowe, David W. ; Lichtler, Alexander C. / 1,25-Dihydroxyvitamin D3 inhibition of Col1a1 promoter expression in calvariae from neonatal transgenic mice. In: BBA - Gene Structure and Expression. 1998 ; Vol. 1398, No. 3. pp. 285-293.
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AU - Bedalov, Antonio

AU - Salvatori, Roberto

AU - Dodig, Milan

AU - Kapural, Belinda

AU - Pavlin, Dubravko

AU - Kream, Barbara E.

AU - Clark, Stephen H.

AU - Woody, Charles O.

AU - Rowe, David W.

AU - Lichtler, Alexander C.

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