Cellular and Molecular Life Sciences

, Volume 68, Issue 23, pp 3885–3901 | Cite as

Roles of the human hypoxia-inducible factor (HIF)-3α variants in the hypoxia response

  • Minna Heikkilä
  • Annika Pasanen
  • Kari I. Kivirikko
  • Johanna Myllyharju
Research article

Abstract

The hypoxia-inducible transcription factor (HIF) controls (in an oxygen-dependent manner) the expression of a large number of genes whose products are involved in the response of cells to hypoxia. HIF is an αβ dimer that binds to hypoxia response elements (HREs) in its target genes. Human HIF-α has three isoforms, HIF-1α, HIF-2α and HIF-3α, of which the roles of HIF-3α are largely unknown, although it is usually regarded as a negative regulator of HIF-1α and HIF-2α. The human HIF-3α locus is subject to extensive alternative splicing, leading to at least seven variants. We analyzed here the effects of the long variants and the short variant HIF-3α4 on the hypoxia response. All these variants were found to interact with HIF-β, HIF-1α and HIF-2α. The long HIF-3α variants were localized in the nucleus in hypoxia, while HIF-3α4 was cytoplasmic. Interaction of the HIF-3α variants with HIF-1α inhibited the nuclear translocation of both. None of the long HIF-3α variants was capable of efficient induction of an HRE reporter in overexpression experiments, but instead inhibited the transcriptional activation of the reporter by HIF-1 and HIF-2. Unexpectedly, siRNA knock-down of the endogenous HIF-3α variants led to downregulation of certain HIF target genes, while overexpression of individual long HIF-3α variants upregulated certain HIF target genes in a variant and target gene-specific manner under conditions in which HIF-β was not a limiting factor. These data indicate that the HIF-3α variants may have more versatile and specific roles in the regulation of the hypoxia response than previously anticipated.

Keywords

Hypoxia response Hypoxia-inducible factor Hypoxia-inducible factor 3 isoform Prolyl 4-hydroxylase Hypoxia response element 

Abbreviations

HIF

Hypoxia-inducible factor

ODDD

Oxygen-dependent degradation domain

P4H

Prolyl 4-hydroxylase

C-TAD

C-terminal transactivation domain

bHLH

Basic helix-loop-helix

PAS

Per-ARNT-Sim

HRE

Hypoxia response element

IPAS

Inhibitory PAS domain protein

LZIP

Leucine zipper

qPCR

Quantitative real-time RT-PCR

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Copyright information

© Springer Basel AG 2011

Authors and Affiliations

  • Minna Heikkilä
    • 1
  • Annika Pasanen
    • 1
  • Kari I. Kivirikko
    • 1
  • Johanna Myllyharju
    • 1
  1. 1.Oulu Center for Cell–Matrix Research, Biocenter Oulu and Department of Medical Biochemistry and Molecular BiologyUniversity of OuluOuluFinland

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