Transcriptional regulation of the major zinc uptake protein hZip1 in prostate cancer cells

Peter Makhov, Konstantin Golovine, Robert G. Uzzo, Torsten Wuestefeld, Benjamin J. Scoll, Vladimir M. Kolenko

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

hZip1 has been characterized as the major zinc uptake transporter regulating the accumulation of zinc in prostate cells. The mechanisms regulating expression of hZip1 have not been described. To explore the mechanisms of transcriptional regulation of the hZip1 gene, we determined the putative promoter sequence for hZip1 and identified the potential transcription start site within the predicted hZip1 promoter region. To further characterize the promoter region for basal hZip1 transcription, 3′ and 5′ deletion constructs and constructs with mutated binding sites for putative transcription factors were generated by PCR amplification and assessed for transcriptional activity with a luciferase reporter assay in PC-3 prostate cancer cells. The ability of the specific transcription factors to bind the hZip1 core promoter was confirmed by EMSA, GelSupershift and ChIP assays. Our experiments identified the core promoter region responsible for constitutive expression of hZip1 and demonstrated critical roles for SP1 and CREB1 in transcriptional regulation of the hZip1 gene in prostate cancer cells.

Original languageEnglish
Pages (from-to)39-46
Number of pages8
JournalGene
Volume431
Issue number1-2
DOIs
StatePublished - Feb 15 2009

Keywords

  • Basal transcription
  • CREB
  • Core promoter
  • SP1

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