Tanimoto K, Makino Y, Pereira T, Poellinger L.. Mechanism of regulation of hypoxia-inducible factor-1 by von Hippel-Lindau tumor suppressor protein. EMBO J 19: 4298-4309

Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institutet, S-171 77 Stockholm, Sweden.
The EMBO Journal (Impact Factor: 10.43). 09/2000; 19(16):4298-309. DOI: 10.1093/emboj/19.16.4298
Source: PubMed


In normoxic cells the hypoxia-inducible factor-1 alpha (HIF-1 alpha) is rapidly degraded by the ubiquitin-proteasome pathway, and activation of HIF-1 alpha to a functional form requires protein stabilization. Here we show that the product of the von Hippel-Lindau (VHL) tumor suppressor gene mediated ubiquitylation and proteasomal degradation of HIF-1 alpha under normoxic conditions via interaction with the core of the oxygen-dependent degradation domain of HIF-1 alpha. The region of VHL mediating interaction with HIF-1 alpha overlapped with a putative macromolecular binding site observed within the crystal structure of VHL. This motif of VHL also represents a mutational hotspot in tumors, and one of these mutations impaired interaction with HIF-1 alpha and subsequent degradation. Interestingly, the VHL binding site within HIF-1 alpha overlapped with one of the minimal transactivation domains. Protection of HIF-1 alpha against degradation by VHL was a multistep mechanism, including hypoxia-induced nuclear translocation of HIF-1 alpha and an intranuclear hypoxia-dependent signal. VHL was not released from HIF-1 alpha during this process. Finally, stabilization of HIF-1 alpha protein levels per se did not totally bypass the need of the hypoxic signal for generating the transactivation response.

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Available from: Teresa Pereira, Sep 30, 2015
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    • "We then determined whether Hif-1a binds to NICD1 in breast cancer cells under hypoxia, as was previously reported (Gustafsson et al., 2005). FLAG-NICD1 and hemagglutinin (HA)-Hif-1a-DODD (a variant of Hif-1a that lacks the oxygen-sensing domain [ODD] and thus is stable in normoxia ; Tanimoto et al., 2000) were coexpressed in MCF-7 cells. However, reciprocal coimmunoprecipitation studies with anti- HA and anti-FLAG failed to reveal any interaction between FLAG-NICD1 and HA-Hif-1a-DODD, while the same antibodies were able to immunoprecipitate their respective targets, Hif- 1a-DODD and NICD1, respectively (Figure 1E). "
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    ABSTRACT: γ-Secretase is composed of four proteins that are obligatory for protease activity: presenilin, nicastrin, Aph1, and Pen-2. Despite the progress toward understanding the function of these individual subunits, there is no information available pertaining to the modulation of γ-secretase in response to environmental changes in cells. Here, we show that hypoxia upregulates γ-secretase activity through a direct interaction with Hif-1α, revealing an unconventional function for Hif-1α as an enzyme subunit, which is distinct from its canonical role as a transcription factor. Moreover, hypoxia-induced cell invasion and metastasis are alleviated by either γ-secretase inhibitors or a dominant-negative Notch coactivator, indicating that γ-secretase/Notch signaling plays an essential role in controlling these cellular processes. The present study reveals a mechanism in which γ-secretase can achieve temporal control through conditional interactions with regulatory proteins, such as Hif-1α, under select physiological and pathological conditions.
    Cell Reports 08/2014; 8(4). DOI:10.1016/j.celrep.2014.07.028 · 8.36 Impact Factor
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    • "The basic biology underlying the development of clear cell renal cell carcinoma is critically dependent on the VHL gene, whose protein product is important in the cell's normal response to hypoxia. Aberrations in VHL's function, either through mutation or promoter hypermethylation, lead to accumulation of the transcriptional regulatory molecule, HIF-α.[56] In view of the importance of VHL gene in such conditions the present study is aimed to characterize VHL in an Indian patient reported in Sri Venkateswara Institute of Medical Sciences (SVIMS), Tirupati in the Department of Urology, with clear cell renal carcinoma as identified in the Department of Pathology. "
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    ABSTRACT: Von Hippel-Lindau (VHL) disease is an autosomal dominant hereditary cancer syndrome that predisposes to the development of a variety of benign and malignant tumors, especially cerebellar hemangioblastomas, retinal angiomas and clear-cell renal cell carcinomas (RCC). We have identified of VHL gene using immunohistochemistry in a patient who was diagnosed for RCC. In order to understand the involvement of mutation in the VHL gene exon 1 was amplified and sequenced (accession number: JX 401534). The sequence analysis revealed the presence of novel missense mutations c.194 C>T, c.239 G>A, c.278 G>A, c.319 C>G, c. 337 C > G leading to the following variations p.Ala 65 Val, p.Gly 80 Asp, p.Gly 93 Glu, p.Gln 107 Glu, p.Gln 113 Glu in the protein.
    Indian Journal of Human Genetics 07/2013; 19(3):373-6. DOI:10.4103/0971-6866.120809
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    • "The PHD proteins are dioxygenases which require oxygen for their function and as such are sensitive to oxygen concentrations, losing their activity under low oxygen concentration [22]. The hydroxylated HIFα proteins are poly-ubiquitinylated and targeted for 26S proteosomal degradation through the von Hippel-Lindau (pVHL)/Elongin BC/Cul2 ubiquitin-ligase complex [28], [29], [30], [31], [32], [33], [34]. Under low oxygen conditions, the PHD proteins are inactive, so the HIFα proteins are not hydroxylated and stable. "
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    ABSTRACT: Lung development occurs under relative hypoxia and the most important oxygen-sensitive response pathway is driven by Hypoxia Inducible Factors (HIF). HIFs are heterodimeric transcription factors of an oxygen-sensitive subunit, HIFα, and a constitutively expressed subunit, HIF1β. HIF1α and HIF2α, encoded by two separate genes, contribute to the activation of hypoxia inducible genes. A third HIFα gene, , is subject to alternative promoter usage and splicing, leading to three major isoforms, HIF3α, NEPAS and IPAS. HIF3α gene products add to the complexity of the hypoxia response as they function as dominant negative inhibitors (IPAS) or weak transcriptional activators (HIF3α/NEPAS). Previously, we and others have shown the importance of the Hif1α and Hif2α factors in lung development, and here we investigated the role of Hif3α during pulmonary development. Therefore, HIF3α was conditionally expressed in airway epithelial cells during gestation and although HIF3α transgenic mice were born alive and appeared normal, their lungs showed clear abnormalities, including a post-pseudoglandular branching defect and a decreased number of alveoli. The HIF3α expressing lungs displayed reduced numbers of Clara cells, alveolar epithelial type I and type II cells. As a result of HIF3α expression, the level of Hif2α was reduced, but that of Hif1α was not affected. Two regulatory genes, Rarβ, involved in alveologenesis, and Foxp2, a transcriptional repressor of the Clara cell specific Ccsp gene, were significantly upregulated in the HIF3α expressing lungs. In addition, aberrant basal cells were observed distally as determined by the expression of Sox2 and p63. We show that Hif3α binds a conserved HRE site in the Sox2 promoter and weakly transactivated a reporter construct containing the Sox2 promoter region. Moreover, Hif3α affected the expression of genes not typically involved in the hypoxia response, providing evidence for a novel function of Hif3α beyond the hypoxia response.
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