Copper levels affect targeting of hypoxia-inducible factor 1 to the promoters of hypoxia-regulated genes

Xiaojuan Liu, Wenjing Zhang, Zhijuan Wu, Yutao Yang, Yujian Kang

Research output: Contribution to journalArticle

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Abstract

Hypoxia-inducible factor 1α (HIF-1α) is a transcription factor that regulates cellular responses to hypoxia. It controls the expression of both BCL2/adenovirus E1B 19-kDa protein–interacting protein 3 (BNIP3) and insulin-like growth factor 2 (IGF2). Previous studies have demonstrated that in hypoxia, copper is required for the expression of BNIP3 but not for that of IGF2. Here, using ChIP assays, computational analyses, luciferase reporter assays, and real-time quantitative RT-PCR, we sought to better understand how copper regulates the differential target gene selectivity of HIF-1α. Human umbilical vein endothelial cells (HUVECs) were exposed to CoCl 2 or hypoxia conditions to increase HIF-1α accumulation. The binding of HIF-1α to hypoxia-responsive element (HRE) sites in the BNIP3 or IGF2 gene promoter in high- or low-copper conditions was examined. Our analyses revealed three and two potential HRE sites in the BNIP3 and IGF2 promoters, respectively. We identified that HRE (-412/-404) in the BNIP3 promoter and HRE (-354/-347) in the IGF2 promoter are the critical binding sites of HIF-1α. Tetraethelenepentamine (TEPA)-mediated reduction in copper concentration did not affect hypoxia- or CoCl 2 -induced HIF-1α accumulation. However, the copper reduction did suppress the binding of HIF-1α to the HRE (-412/-404) in BNIP3 but not the binding of HIF-1α to the HRE (-354/-347) in IGF2. In summary, our findings uncovered the mechanistic basis for differential HIF-1α–mediated regulation of BNIP3 and IGF2, indicating that copper regulates target gene selectivity of HIF-1α at least in part by affecting HIF-1α binding to its cognate HRE in the promoters of these two genes.

Original languageEnglish (US)
Pages (from-to)14669-14677
Number of pages9
JournalJournal of Biological Chemistry
Volume293
Issue number38
DOIs
StatePublished - Jan 1 2018

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Hypoxia-Inducible Factor 1
Copper
Genes
Somatomedins
Assays
Hypoxia
Endothelial cells
Luciferases
Human Umbilical Vein Endothelial Cells
Adenoviridae
Transcription Factors
Real-Time Polymerase Chain Reaction
Binding Sites

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Copper levels affect targeting of hypoxia-inducible factor 1 to the promoters of hypoxia-regulated genes. / Liu, Xiaojuan; Zhang, Wenjing; Wu, Zhijuan; Yang, Yutao; Kang, Yujian.

In: Journal of Biological Chemistry, Vol. 293, No. 38, 01.01.2018, p. 14669-14677.

Research output: Contribution to journalArticle

Liu, Xiaojuan ; Zhang, Wenjing ; Wu, Zhijuan ; Yang, Yutao ; Kang, Yujian. / Copper levels affect targeting of hypoxia-inducible factor 1 to the promoters of hypoxia-regulated genes. In: Journal of Biological Chemistry. 2018 ; Vol. 293, No. 38. pp. 14669-14677.
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abstract = "Hypoxia-inducible factor 1α (HIF-1α) is a transcription factor that regulates cellular responses to hypoxia. It controls the expression of both BCL2/adenovirus E1B 19-kDa protein–interacting protein 3 (BNIP3) and insulin-like growth factor 2 (IGF2). Previous studies have demonstrated that in hypoxia, copper is required for the expression of BNIP3 but not for that of IGF2. Here, using ChIP assays, computational analyses, luciferase reporter assays, and real-time quantitative RT-PCR, we sought to better understand how copper regulates the differential target gene selectivity of HIF-1α. Human umbilical vein endothelial cells (HUVECs) were exposed to CoCl 2 or hypoxia conditions to increase HIF-1α accumulation. The binding of HIF-1α to hypoxia-responsive element (HRE) sites in the BNIP3 or IGF2 gene promoter in high- or low-copper conditions was examined. Our analyses revealed three and two potential HRE sites in the BNIP3 and IGF2 promoters, respectively. We identified that HRE (-412/-404) in the BNIP3 promoter and HRE (-354/-347) in the IGF2 promoter are the critical binding sites of HIF-1α. Tetraethelenepentamine (TEPA)-mediated reduction in copper concentration did not affect hypoxia- or CoCl 2 -induced HIF-1α accumulation. However, the copper reduction did suppress the binding of HIF-1α to the HRE (-412/-404) in BNIP3 but not the binding of HIF-1α to the HRE (-354/-347) in IGF2. In summary, our findings uncovered the mechanistic basis for differential HIF-1α–mediated regulation of BNIP3 and IGF2, indicating that copper regulates target gene selectivity of HIF-1α at least in part by affecting HIF-1α binding to its cognate HRE in the promoters of these two genes.",
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AU - Kang, Yujian

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AB - Hypoxia-inducible factor 1α (HIF-1α) is a transcription factor that regulates cellular responses to hypoxia. It controls the expression of both BCL2/adenovirus E1B 19-kDa protein–interacting protein 3 (BNIP3) and insulin-like growth factor 2 (IGF2). Previous studies have demonstrated that in hypoxia, copper is required for the expression of BNIP3 but not for that of IGF2. Here, using ChIP assays, computational analyses, luciferase reporter assays, and real-time quantitative RT-PCR, we sought to better understand how copper regulates the differential target gene selectivity of HIF-1α. Human umbilical vein endothelial cells (HUVECs) were exposed to CoCl 2 or hypoxia conditions to increase HIF-1α accumulation. The binding of HIF-1α to hypoxia-responsive element (HRE) sites in the BNIP3 or IGF2 gene promoter in high- or low-copper conditions was examined. Our analyses revealed three and two potential HRE sites in the BNIP3 and IGF2 promoters, respectively. We identified that HRE (-412/-404) in the BNIP3 promoter and HRE (-354/-347) in the IGF2 promoter are the critical binding sites of HIF-1α. Tetraethelenepentamine (TEPA)-mediated reduction in copper concentration did not affect hypoxia- or CoCl 2 -induced HIF-1α accumulation. However, the copper reduction did suppress the binding of HIF-1α to the HRE (-412/-404) in BNIP3 but not the binding of HIF-1α to the HRE (-354/-347) in IGF2. In summary, our findings uncovered the mechanistic basis for differential HIF-1α–mediated regulation of BNIP3 and IGF2, indicating that copper regulates target gene selectivity of HIF-1α at least in part by affecting HIF-1α binding to its cognate HRE in the promoters of these two genes.

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