mRNA cloning and expression studies of the quail homologue of HIF-2α

Elvert, Gerd; Lanz, Stephan; Kappel, Andreas; Flamme, Ingo

doi:10.1016/s0925-4773(99)00144-6

Cited by 14 publications

(17 citation statements)

References 13 publications

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“…Shifting from oxidative phosphorylation to the O 2 -independent glycolytic pathway is the most ancient cellular response to hypoxia, occurring even in Caenorhabditis elegans (77). Interestingly, HIF-1␣ is conserved from C. elegans to humans and HIF-2␣ only exists in more complicated vertebrates such as chicken, quail, and mammals (24,28,47). Involvement of HIF-1␣, and not HIF-2␣, in this ancient glycolytic pathway makes sense based on the fact that HIF-1␣ is evolutionarily conserved and ubiquitously expressed.…”

Section: Discussionmentioning

confidence: 99%

“…Northern blot analysis was performed according to standard protocols (36). Human DNA fragments composed of the indicated nucleotides (nt) (with section numbers of the genes from GenBank) were generated through reverse transcription-PCR and cloned into pCRII-topo vector to serve as templates for 24 primers. Subsequently, biotin-labeled ribonucleotides were synthesized with a BioArray high-yield RNA transcript labeling kit (Enzo Diagnostic, Inc.).…”

Section: Methodsmentioning

confidence: 99%

“…1, lanes 1 to 4), a result that is consistent with previous reports (66). Importantly, no HIF-1␣ protein was detected even in the VHL-defective 786-O PRC-3 cells ( To systematically study HIF-2␣-inducible genes, triplicate RNA samples were prepared from 786-O WT-8 cells cultured under normoxia (21% O 2 ) or hypoxia (1.5% O 2 ) for 16 h. Double-stranded cDNA was generated from total RNA with the Superscript Choice System (Gibco BRL Life Technologies) and T7-(dT) 24 primers. Subsequently, antisense biotinlabeled RNA was synthesized with T7 DNA polymerase.…”

Section: Hypoxia-inducible Genes In 786-o Wt-8 Cells Both Hif-1␣mentioning

confidence: 99%

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Differential Roles of Hypoxia-Inducible Factor 1α (HIF-1α) and HIF-2α in Hypoxic Gene Regulation

Wang

Chodosh

et al. 2003

Molecular and Cellular Biology

1,236

1,078

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Transcriptional responses to hypoxia are primarily mediated by hypoxia-inducible factor (HIF), a heterodimer of HIF-␣ and the aryl hydrocarbon receptor nuclear translocator subunits. The HIF-1␣ and HIF-2␣ subunits are structurally similar in their DNA binding and dimerization domains but differ in their transactivation domains, implying they may have unique target genes. Previous studies using Hif-1␣ ؊/؊ embryonic stem and mouse embryonic fibroblast cells show that loss of HIF-1␣ eliminates all oxygen-regulated transcriptional responses analyzed, suggesting that HIF-2␣ is dispensable for hypoxic gene regulation. In contrast, HIF-2␣ has been shown to regulate some hypoxia-inducible genes in transient transfection assays and during embryonic development in the lung and other tissues. To address this discrepancy, and to identify specific HIF-2␣ target genes, we used DNA microarray analysis to evaluate hypoxic gene induction in cells expressing HIF-2␣ but not HIF-1␣. In addition, we engineered HEK293 cells to express stabilized forms of HIF-1␣ or HIF-2␣ via a tetracycline-regulated promoter. In this first comparative study of HIF-1␣ and HIF-2␣ target genes, we demonstrate that HIF-2␣ does regulate a variety of broadly expressed hypoxia-inducible genes, suggesting that its function is not restricted, as initially thought, to endothelial cell-specific gene expression. Importantly, HIF-1␣ (and not HIF-2␣) stimulates glycolytic gene expression in both types of cells, clearly showing for the first time that HIF-1␣ and HIF-2␣ have unique targets.Oxygen (O 2 ), the final electron acceptor during oxidative phosphorylation, is absolutely required for invertebrate and vertebrate life. The immediate response to O 2 deprivation (hypoxia) is a defense phase, which suppresses ATP consumption by arresting protein translation and ion channel activity, two major ATP sinks during normoxia. During a rescue phase, in spite of a general reduction in RNA synthesis, transcription of some genes increases dramatically under low O 2 (21, 34). These hypoxia-responsive genes are involved in glucose transport, glycolysis, erythropoiesis, angiogenesis, vasodilation, and respiratory rate, and together they function to minimize the effects caused by low O 2 at cellular, tissue and systemic levels (93, 106).The activation of many O 2 -regulated genes is mediated by hypoxia-inducible factor (HIF), a heterodimer consisting of HIF-1␣ and HIF-1␤ (also called the aryl hydrocarbon receptor nuclear translocator [ARNT]) in most cells (52,104,105). Both HIF-1␣ and ARNT belong to the basic helix-loop-helix (bHLH)-Per-Arnt-Sim (PAS) family of transcription factors, which share several conserved structural domains, including a bHLH region for DNA binding and two PAS domains for target gene specificity and dimerization (102). Although ARNT is absolutely required for HIF activity (63, 110), HIF function is primarily regulated by HIF-1␣ protein stability (37,46,84). Under normoxia, HIF-1␣ is ubiquitinated through interaction with the von Hippel-Lindau tumor suppr...

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Hypoxia-inducible Genes In 786-o Wt-8 Cells Both Hif-1␣mentioning

confidence: 99%

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Differential Roles of Hypoxia-Inducible Factor 1α (HIF-1α) and HIF-2α in Hypoxic Gene Regulation

Wang

Chodosh

et al. 2003

Molecular and Cellular Biology

1,236

1,078

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“…This might reflect their distinct expression patterns and functions. For example, HIF-1␣ is present in organisms from Caenorhabditis elegans to humans, while HIF-2␣ exists only in more complicated vertebrate species, such as chicken, quail, and mammals (9,10,20). In addition, HIF-1␣ is universally expressed while HIF-2␣ expression is more tissue restricted, even in organisms where both isoforms are expressed.…”

Section: Discussionmentioning

confidence: 99%

Differential Regulation of the Transcriptional Activities of Hypoxia-Inducible Factor 1 Alpha (HIF-1α) and HIF-2α in Stem Cells

Iyer

Sataur

et al. 2006

Molecular and Cellular Biology

251

236

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“…Gene targeting experiments have clearly demonstrated that HIF1␣ is required for embryonic vascularization, as major defects in early vascular de-velopment were observed in homozygous mice lacking HIF1␣ (Iyer et al, 1998, Ryan et al, 1998. EPAS1 displays a more restricted expression pattern (Flamme et al, 1997, Jain et al, 1998, Elvert et al, 1999, Favier et al, 1999 and is detected mostly in the vasculature during embryonic development. In vitro studies have shown that it can transactivate the synthesis of VEGF (Ema et al, 1997), its receptor VEGFR-2 , and the angiopoietin receptor Tie2 (Tian et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Coexpression of endothelial PAS protein 1 with essential angiogenic factors suggests its involvement in human vascular development

Favier

Kempf

Corvol

et al. 2001

Developmental Dynamics

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Endothelial PAS protein 1 (EPAS1) is a bHLH-PAS transcription factor involved in cellular response to hypoxia. Its precise role in angiogenesis is unclear, but several genes essential to vascular development, including those encoding vascular endothelial growth factor (VEGF), its receptor VEGFR-2 and Tie2, are thought to be targets of EPAS1. To investigate whether this transcription factor and its putative targets were expressed concomitantly, we performed in situ hybridization on serial adjacent sections of human embryos at gestational ages of 3 to 6 weeks. We studied expression of the genes encoding EPAS1, VEGF, VEGFR-1, and -2, Tie2, and its ligands, angiopoietin (Ang) 1 and 2. We also compared these expression profiles with that of hypoxia-inducible factor 1␣ (HIF1␣). EPAS1 transcripts were detected in several types of endothelial cell: in blood vessels walls, the endocardium, the glomeruli of the mesonephros, and the sinusoids of the liver. In these endothelial cells, expression of EPAS1 systematically or partly coincided with Tie2 and the VEGF receptors expression. There was also some overlap between the sites of synthesis of EPAS1 and VEGF mRNAs, principally in hepatocytes and sympathetic ganglion cells. In addition, we found that EPAS1 and HIF1␣ transcripts were often colocalized, suggesting a functional redundancy of these two transcription factors during development. These observations are consistent with transactivation by EPAS1 of the expression of its putative target genes during embryogenesis, suggesting that this transcription factor is involved in human angiogenesis. They provide evidence that EPAS1 is involved in the regulation of vascular maturation, remodeling, or stabilization rather than in the early steps of embryonic angiogenesis.

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mRNA cloning and expression studies of the quail homologue of HIF-2α

Cited by 14 publications

References 13 publications

Differential Roles of Hypoxia-Inducible Factor 1α (HIF-1α) and HIF-2α in Hypoxic Gene Regulation

Differential Roles of Hypoxia-Inducible Factor 1α (HIF-1α) and HIF-2α in Hypoxic Gene Regulation

Differential Regulation of the Transcriptional Activities of Hypoxia-Inducible Factor 1 Alpha (HIF-1α) and HIF-2α in Stem Cells

Coexpression of endothelial PAS protein 1 with essential angiogenic factors suggests its involvement in human vascular development

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