The promoter region of the human PMCA1 gene mediates transcriptional downregulation by 1,25-dihydroxyvitamin D3

Paul Glendenning, Thomas Ratajczak, Richard L. Prince, Nandor Garamszegi, Emanuel E. Strehler

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The gene for plasma membrane calcium pump isoform I (PMCA1) is expressed in calcium-transporting epithelia and bone mesenchymal cells and is upregulated to 1,25-(OH)2D3 in those tissues. A candidate sequence for a vitamin D response element (VDRE) is present within a 1.7-kb promoter region of the human PMCA1 (hPMCA1) gene. We studied hPMCA1 promoter actfvity fn MDBK and ROS 17/2.8 cell lines as PMCA1 mRNA expression is upregulated by 1,25-(OH)2D3 in both. Structural analysis of the putative hPMCA1 VDRE sequence was performed using mobility shift analysis (EMSA) and nuclear extracts from COS-1 cells expressing human VDR (hVDR) and RXRα (hRXRα). 1,25-(OH)2D3 induced transrepression of the entire 1.7-kb hPMCA1 promoter and of one promoter deletion construct in ROS 17/2.8 cells but not MDBK cells when assayed by luciferase reporter gene assays. Three additional hPMCA1 promoter deletion constructs were unaffected by 1,25-(OH)2D3 in either cell line. While hVDR and hRXRα were capable of complexing with a rat osteocalcin DR3 VDRE, EMSA analysis of the potential VDRE from the hPMCA1 gene did not show interaction of either nuclear receptor. Our results indicate tissue-specific sensitivity of the promoter region of the hPMCA1 gene to direct transcriptional downregulation by 1,25-(OH)2D3 and suggest that any positive regulatory VDRE must lie outside of the 1.7-kb core promoter. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)722-728
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume277
Issue number3
DOIs
StatePublished - Nov 2 2000

Fingerprint

Vitamin D Response Element
Calcitriol
Genetic Promoter Regions
Down-Regulation
Genes
Cells
Tissue
Calcium
Osteocalcin
Cell membranes
Cytoplasmic and Nuclear Receptors
Luciferases
Structural analysis
Rats
Assays
Protein Isoforms
Bone
Pumps
Cell Line
COS Cells

Keywords

  • Calcitriol
  • Calcium transport
  • Kidney distal tubule
  • Osteoblast
  • Plasma membrane calcium ATPase
  • Promoter

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

The promoter region of the human PMCA1 gene mediates transcriptional downregulation by 1,25-dihydroxyvitamin D3 . / Glendenning, Paul; Ratajczak, Thomas; Prince, Richard L.; Garamszegi, Nandor; Strehler, Emanuel E.

In: Biochemical and Biophysical Research Communications, Vol. 277, No. 3, 02.11.2000, p. 722-728.

Research output: Contribution to journalArticle

Glendenning, Paul ; Ratajczak, Thomas ; Prince, Richard L. ; Garamszegi, Nandor ; Strehler, Emanuel E. / The promoter region of the human PMCA1 gene mediates transcriptional downregulation by 1,25-dihydroxyvitamin D3 In: Biochemical and Biophysical Research Communications. 2000 ; Vol. 277, No. 3. pp. 722-728.
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AB - The gene for plasma membrane calcium pump isoform I (PMCA1) is expressed in calcium-transporting epithelia and bone mesenchymal cells and is upregulated to 1,25-(OH)2D3 in those tissues. A candidate sequence for a vitamin D response element (VDRE) is present within a 1.7-kb promoter region of the human PMCA1 (hPMCA1) gene. We studied hPMCA1 promoter actfvity fn MDBK and ROS 17/2.8 cell lines as PMCA1 mRNA expression is upregulated by 1,25-(OH)2D3 in both. Structural analysis of the putative hPMCA1 VDRE sequence was performed using mobility shift analysis (EMSA) and nuclear extracts from COS-1 cells expressing human VDR (hVDR) and RXRα (hRXRα). 1,25-(OH)2D3 induced transrepression of the entire 1.7-kb hPMCA1 promoter and of one promoter deletion construct in ROS 17/2.8 cells but not MDBK cells when assayed by luciferase reporter gene assays. Three additional hPMCA1 promoter deletion constructs were unaffected by 1,25-(OH)2D3 in either cell line. While hVDR and hRXRα were capable of complexing with a rat osteocalcin DR3 VDRE, EMSA analysis of the potential VDRE from the hPMCA1 gene did not show interaction of either nuclear receptor. Our results indicate tissue-specific sensitivity of the promoter region of the hPMCA1 gene to direct transcriptional downregulation by 1,25-(OH)2D3 and suggest that any positive regulatory VDRE must lie outside of the 1.7-kb core promoter. (C) 2000 Academic Press.

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KW - Osteoblast

KW - Plasma membrane calcium ATPase

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