Two Vitamin D Response Elements Function in the Rat 1,25-Dihydroxyvitamin D 24-Hydroxylase Promoter (∗)

The interaction between the two vitamin D response elements (DRE) located at −154 to −134 base pairs (bp) and −262 to −238 bp from the transcription initiation site has been studied using reporter gene assays and binding assays by electrophoretic gel shift measurements. 3 half-sites separated by 3 b...

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Veröffentlicht in:The Journal of biological chemistry 1995-01, Vol.270 (4), p.1675-1678
Hauptverfasser: Zierold, Claudia, Darwish, Hisham M., DeLuca, Hector F.
Format: Artikel
Sprache:eng
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Zusammenfassung:The interaction between the two vitamin D response elements (DRE) located at −154 to −134 base pairs (bp) and −262 to −238 bp from the transcription initiation site has been studied using reporter gene assays and binding assays by electrophoretic gel shift measurements. 3 half-sites separated by 3 bp were found necessary for transactivation by the −154 to −125 DRE, while 2 half-sites separated by 3 bp were needed for the DRE at −262 to −238 to function. However, the two DREs together provided maximal activity. The 93-bp fragment separating the two DREs was not required and could be deleted. The most effective binding by receptor was found with the two complete DREs (dissociation constant (Kd) = 13.7 pM), although each DRE bound to the receptor and nuclear accessory factor with about 5 nMKd. The two DREs (a total of 5 half-sites) apparently account for most if not all of the transactivation of the rat 24-hydroxylase by 1,25-dihydroxyvitamin D3. This system represents the most powerful of the DREs reported to date.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.270.4.1675