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Nucleic Acids Research 2005 33(17):5521-5532; doi:10.1093/nar/gki872
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Published online 26 September 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oxfordjournals.org

Article

Regulation of multiple insulin-like growth factor binding protein genes by 1{alpha},25-dihydroxyvitamin D3

Merja Matilainen, Marjo Malinen, Katri Saavalainen and Carsten Carlberg*

Department of Biochemistry, University of Kuopio P.O. box 1627 IN-70211 Kuopio, Finland

*To whom correspondence should be addressed. Tel: +358 17 163062; Fax: +358 17 2811510; Email: carlberg{at}messi.uku.fi

Received June 8, 2005. Revised September 12, 2005. Accepted September 12, 2005.

Recently, insulin-like growth factor binding proteins (IGFBPs) have been found to be primary mediators of the anti-proliferative actions of the nuclear hormone 1{alpha},25-dihydroxyvitamin D3 [1{alpha},25(OH)2D3], but dependent on cellular context IGFBPs can also have a mitogenic effect. In this study, we performed expression profiling of all six human IGFBP genes in prostate and bone cancer cells and demonstrated that IGFBP1, 3 and 5 are primary 1{alpha},25(OH)2D3 target genes. In silico screening of the 174 kb of genomic sequence surrounding all six IGFBP genes identified 15 candidate vitamin D response elements (VDREs) close to or in IGFBP1, 2, 3 and 5 but not in the IGFBP4 and 6 genes. The putative VDREs were evaluated in vitro by gelshift assays and in living cells by reporter gene and chromatin immuno-precipitation (ChIP) assays. Of these 10 VDREs appear to be functional. ChIP assays demonstrated for each of these an individual, stimulation time-dependent association profile not only with the vitamin D receptor, but also with first heterodimeric partner the retinoid X receptor, other regulatory complex components and phosphorylated RNA polymerase II. Some of the VDREs are located distantly from the transcription start sites of IGFBP1, 3 and 5, but all 10 VDREs seem to contribute to the regulation of the genes by 1{alpha},25(OH)2D3. In conclusion, IGFBP1, 3 and 5 are primary 1{alpha},25(OH)2D3 target genes that in intact cells are each under the control of multiple VDREs.


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