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1,25-Dihydroxyvitamin D3 inhibition of col1a1 promoter expression in calvariae from neonatal transgenic miceWe 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.
Document ID
20040172638
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Bedalov, A.
(University of Connecticut Health Center 263 Farmington Ave., Farmington, CT 06030, United States)
Salvatori, R.
Dodig, M.
Kapural, B.
Pavlin, D.
Kream, B. E.
Clark, S. H.
Woody, C. O.
Rowe, D. W.
Lichtler, A. C.
Date Acquired
August 22, 2013
Publication Date
July 9, 1998
Publication Information
Publication: Biochimica et biophysica acta
Volume: 1398
Issue: 3
ISSN: 0006-3002
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: AR38933
CONTRACT_GRANT: AR2985
CONTRACT_GRANT: AR29983
Distribution Limits
Public
Copyright
Other
Keywords
NASA Discipline Cell Biology
Non-NASA Center

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