Cell-specific regulation of hypoxia-inducible factor (HIF)-1α and HIF-2α stabilization and transactivation in a graded oxygen environment

Cameron P. Bracken, Anthony O. Fedele, Sarah Linke, Wiltiana Balrak, Karolina Lisy, Murray L. Whitelaw, Daniel J. Peet

Research output: Contribution to journalArticle

120 Citations (Scopus)

Abstract

The hypoxia-inducible factor (HIF)-1α and HIF-2α are closely related, key transcriptional regulators of the hypoxic response, countering a low oxygen situation with the up-regulation of target genes associated with numerous processes, including vascularization and glycolysis. This involves a dual mechanism of control through both stabilization and transactivation, regulated via prolyl and asparaginyl hydroxylation. Despite high similarity with respect to protein sequence and activation pathway, a growing number of physiological and mechanistic differences between HIF-1α and HIF-2α are being reported. To further characterize this nonredundancy, the stabilization of endogenous proteins and regulation of the transactivation domains were compared in a graded oxygen environment across a series of cell lines. Although generally similar results were found, interesting and specific differences between the HIF-α proteins were observed within certain cell lines, such as rat adrenal PC12s, emphasizing the cell-specific nature of HIF-α regulation. We characterize a conserved amino acid substitution between HIF-1α and HIF-2α that contributes to the intrinsically higher FIH-1-mediated asparaginyl hydroxylation of HIF-1α and, hence, lower HIF-1α activity. In addition, our data demonstrate that the different cell lines can be classified into two distinct groups: those in which stabilization and transactivation proceed in conjunction (HeLa, 293T, and COS-1) and those cells in which HIF-α is stabilized prior to transactivation (PC12, HepG2, and CACO2). Interestingly, the initial stabilization of HIF-α prior to transactivation up-regulation predicted from in vitro derived hydroxylation data is only true for a subset of cells.

LanguageEnglish
Pages22575-22585
Number of pages11
JournalJournal of Biological Chemistry
Volume281
Issue number32
DOIs
Publication statusPublished - 11 Aug 2006
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Bracken, Cameron P. ; Fedele, Anthony O. ; Linke, Sarah ; Balrak, Wiltiana ; Lisy, Karolina ; Whitelaw, Murray L. ; Peet, Daniel J. / Cell-specific regulation of hypoxia-inducible factor (HIF)-1α and HIF-2α stabilization and transactivation in a graded oxygen environment. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 32. pp. 22575-22585.
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Cell-specific regulation of hypoxia-inducible factor (HIF)-1α and HIF-2α stabilization and transactivation in a graded oxygen environment. / Bracken, Cameron P.; Fedele, Anthony O.; Linke, Sarah; Balrak, Wiltiana; Lisy, Karolina; Whitelaw, Murray L.; Peet, Daniel J.

In: Journal of Biological Chemistry, Vol. 281, No. 32, 11.08.2006, p. 22575-22585.

Research output: Contribution to journalArticle

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T1 - Cell-specific regulation of hypoxia-inducible factor (HIF)-1α and HIF-2α stabilization and transactivation in a graded oxygen environment

AU - Bracken, Cameron P.

AU - Fedele, Anthony O.

AU - Linke, Sarah

AU - Balrak, Wiltiana

AU - Lisy, Karolina

AU - Whitelaw, Murray L.

AU - Peet, Daniel J.

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N2 - The hypoxia-inducible factor (HIF)-1α and HIF-2α are closely related, key transcriptional regulators of the hypoxic response, countering a low oxygen situation with the up-regulation of target genes associated with numerous processes, including vascularization and glycolysis. This involves a dual mechanism of control through both stabilization and transactivation, regulated via prolyl and asparaginyl hydroxylation. Despite high similarity with respect to protein sequence and activation pathway, a growing number of physiological and mechanistic differences between HIF-1α and HIF-2α are being reported. To further characterize this nonredundancy, the stabilization of endogenous proteins and regulation of the transactivation domains were compared in a graded oxygen environment across a series of cell lines. Although generally similar results were found, interesting and specific differences between the HIF-α proteins were observed within certain cell lines, such as rat adrenal PC12s, emphasizing the cell-specific nature of HIF-α regulation. We characterize a conserved amino acid substitution between HIF-1α and HIF-2α that contributes to the intrinsically higher FIH-1-mediated asparaginyl hydroxylation of HIF-1α and, hence, lower HIF-1α activity. In addition, our data demonstrate that the different cell lines can be classified into two distinct groups: those in which stabilization and transactivation proceed in conjunction (HeLa, 293T, and COS-1) and those cells in which HIF-α is stabilized prior to transactivation (PC12, HepG2, and CACO2). Interestingly, the initial stabilization of HIF-α prior to transactivation up-regulation predicted from in vitro derived hydroxylation data is only true for a subset of cells.

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