Mechanism of regulation of the hypoxia‐inducible factor‐1α by the von Hippel‐Lindau tumor suppressor protein

In normoxic cells the hypoxia‐inducible factor‐1α (HIF‐1α) is rapidly degraded by the ubiquitin‐proteasome pathway, and activation of HIF‐1α 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‐1α under normoxic conditions via interaction with the core of the oxygen‐dependent degradation domain of HIF‐1α. The region of VHL mediating interaction with HIF‐1α 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‐1α and subsequent degradation. Interestingly, the VHL binding site within HIF‐1α overlapped with one of the minimal transactivation domains. Protection of HIF‐1α against degradation by VHL was a multistep mechanism, including hypoxia‐induced nuclear translocation of HIF‐1α and an intranuclear hypoxia‐dependent signal. VHL was not released from HIF‐1α during this process. Finally, stabilization of HIF‐1α protein levels per se did not totally bypass the need of the hypoxic signal for generating the transactivation response.

[1]  G. Semenza,et al.  Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[2]  W. Kaelin,et al.  Negative regulation of hypoxia-inducible genes by the von Hippel-Lindau protein. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[3]  R. Klausner,et al.  Post-transcriptional regulation of vascular endothelial growth factor mRNA by the product of the VHL tumor suppressor gene. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. Gassmann,et al.  Functional interference between hypoxia and dioxin signal transduction pathways: competition for recruitment of the Arnt transcription factor , 1996, Molecular and cellular biology.

[5]  S. Bhattacharya,et al.  An essential role for p300/CBP in the cellular response to hypoxia. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[6]  L. Poellinger,et al.  Activation of hypoxia-inducible factor 1alpha: posttranscriptional regulation and conformational change by recruitment of the Arnt transcription factor. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. A. Atarinagradin.  Activation of hypoxia-inducible factor 1a: Posttranscriptional regulation and conformational change by recruitment of the Arnt transcription factor , 1997 .

[8]  G. Semenza,et al.  Transactivation and Inhibitory Domains of Hypoxia-inducible Factor 1α , 1997, The Journal of Biological Chemistry.

[9]  J. Caro,et al.  Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes. , 1997, The Journal of biological chemistry.

[10]  M. Gassmann,et al.  Oxygen(es) and the hypoxia-inducible factor-1. , 1997, Biological chemistry.

[11]  M. Nagao,et al.  Activation of Hypoxia-inducible Factor-1; Definition of Regulatory Domains within the α Subunit* , 1997, The Journal of Biological Chemistry.

[12]  D. Mukhopadhyay,et al.  The von Hippel-Lindau tumor suppressor gene product interacts with Sp1 to repress vascular endothelial growth factor promoter activity , 1997, Molecular and cellular biology.

[13]  M. Gassmann,et al.  Cellular and developmental control of O2 homeostasis by hypoxia-inducible factor 1 alpha. , 1998, Genes & development.

[14]  L. Poellinger,et al.  Signal transduction in hypoxic cells: inducible nuclear translocation and recruitment of theCBP/p300 coactivator by the hypoxia‐induciblefactor‐1α , 1998, The EMBO journal.

[15]  Aaron Ciechanover,et al.  The ubiquitin–proteasome pathway: on protein death and cell life , 1998, The EMBO journal.

[16]  W. Kaelin,et al.  The VHL tumour-suppressor gene paradigm. , 1998, Trends in genetics : TIG.

[17]  W. Kaelin,et al.  Regulation of Hypoxia-Inducible mRNAs by the von Hippel-Lindau Tumor Suppressor Protein Requires Binding to Complexes Containing Elongins B/C and Cul2 , 1998, Molecular and Cellular Biology.

[18]  L. Huang,et al.  Regulation of hypoxia-inducible factor 1α is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway , 1998 .

[19]  C. Wykoff,et al.  The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis , 1999, Nature.

[20]  R. Klausner,et al.  Transcription-Dependent Nuclear-Cytoplasmic Trafficking Is Required for the Function of the von Hippel-Lindau Tumor Suppressor Protein , 1999, Molecular and Cellular Biology.

[21]  I. Zhulin,et al.  PAS Domains: Internal Sensors of Oxygen, Redox Potential, and Light , 1999, Microbiology and Molecular Biology Reviews.

[22]  C. Thrash-Bingham,et al.  aHIF: a natural antisense transcript overexpressed in human renal cancer and during hypoxia. , 1999, Journal of the National Cancer Institute.

[23]  M. Hochstrasser,et al.  Substrate Targeting in the Ubiquitin System , 1999, Cell.

[24]  Y. Fujii‐Kuriyama,et al.  Molecular mechanisms of transcription activation by HLF and HIF1α in response to hypoxia: their stabilization and redox signal‐induced interaction with CBP/p300 , 1999, The EMBO journal.

[25]  J. Caro,et al.  Characterization of an oxygen/redox-dependent degradation domain of hypoxia-inducible factor alpha (HIF-alpha) proteins. , 1999, Biochemical and biophysical research communications.

[26]  Yuichi Makino,et al.  Regulation of the Hypoxia-inducible Transcription Factor 1α by the Ubiquitin-Proteasome Pathway* , 1999, The Journal of Biological Chemistry.

[27]  R. Klausner,et al.  Identification of the von Hippel-lindau tumor-suppressor protein as part of an active E3 ubiquitin ligase complex. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[28]  M. Gstaiger,et al.  The von Hippel-Lindau tumor suppressor protein is a component of an E3 ubiquitin-protein ligase activity. , 1999, Genes & development.

[29]  W. Kaelin,et al.  Structure of the VHL-ElonginC-ElonginB complex: implications for VHL tumor suppressor function. , 1999, Science.

[30]  L. Poellinger,et al.  Redox-Regulated Recruitment of the Transcriptional Coactivators CREB-Binding Protein and SRC-1 to Hypoxia-Inducible Factor 1α , 2000, Molecular and Cellular Biology.