A mechanism for induction of a hypoxic response by vaccinia virus

Viruses have evolved sophisticated strategies to exploit host cell function for their benefit. Here we show that under physiologically normal oxygen levels (normoxia) vaccinia virus (VACV) infection leads to a rapid stabilization of hypoxia-inducible factor (HIF)-1α, its translocation into the nucleus and the activation of HIF-responsive genes, such as vascular endothelial growth factor (VEGF), glucose transporter-1, and pyruvate dehydrogenase kinase-1. HIF-1α stabilization is mediated by VACV protein C16 that binds the human oxygen sensing enzyme prolyl-hydroxylase domain containing protein (PHD)2 and thereby inhibits PHD2-dependent hydroxylation of HIF-1α. The binding between C16 and PHD2 is direct and specific, and ectopic expression of C16 alone induces transcription of HIF-1α responsive genes. Conversely, a VACV strain lacking the gene for C16, C16L, is unable to induce HIF-1α stabilization. Interestingly, the N-terminal region of C16 is predicted to have a PHD2-like structural fold but lacks the catalytic active site residues of PHDs. The induction of a hypoxic response by VACV is reminiscent of the biochemical consequences of solid tumor formation, and illustrates a poxvirus strategy for manipulation of cellular gene expression and biochemistry.

[1]  S. Kyo,et al.  HYPOXIA-SPECIFIC STABILIZATION OF HIF-1ALPHA BY HUMAN PAPILLOMAVIRSUES , 2009 .

[2]  B. Moss,et al.  Recognition of cloned influenza virus hemagglutinin gene products by cytotoxic T lymphocytes , 1986, Journal of virology.

[3]  C. Lo,et al.  White Spot Syndrome Virus Induces Metabolic Changes Resembling the Warburg Effect in Shrimp Hemocytes in the Early Stage of Infection , 2011, Journal of Virology.

[4]  A. Harris,et al.  Differential Function of the Prolyl Hydroxylases PHD1, PHD2, and PHD3 in the Regulation of Hypoxia-inducible Factor* , 2004, Journal of Biological Chemistry.

[5]  G. Smith,et al.  Vaccinia virus gene A36R encodes a M(r) 43-50 K protein on the surface of extracellular enveloped virus. , 1994, Virology.

[6]  J. Seong,et al.  Hepatitis B Virus X Protein Enhances Transcriptional Activity of Hypoxia-inducible Factor-1α through Activation of Mitogen-activated Protein Kinase Pathway* , 2003, Journal of Biological Chemistry.

[7]  J. Zondlo,et al.  Cellular oxygen sensing: Crystal structure of hypoxia-inducible factor prolyl hydroxylase (PHD2). , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[8]  G. Semenza,et al.  Hypoxia-Inducible Factors in Physiology and Medicine , 2012, Cell.

[9]  S. Kondo,et al.  Epstein-Barr Virus Latent Membrane Protein 1 Induces Synthesis of Hypoxia-Inducible Factor 1α , 2004, Molecular and Cellular Biology.

[10]  S. McFarlane,et al.  Interaction of the human cytomegalovirus particle with the host cell induces hypoxia-inducible factor 1 alpha. , 2011, Virology.

[11]  L. Johnston,et al.  Vaccinia virus encodes an active thymidylate kinase that complements a cdc8 mutant of Saccharomyces cerevisiae. , 1991, The Journal of biological chemistry.

[12]  P. Brown,et al.  Stunned Silence: Gene Expression Programs in Human Cells Infected with Monkeypox or Vaccinia Virus , 2011, PloS one.

[13]  N. Roseman,et al.  Vaccinia virus-encoded ribonucleotide reductase: sequence conservation of the gene for the small subunit and its amplification in hydroxyurea-resistant mutants , 1988, Journal of virology.

[14]  C. Boshoff,et al.  Identification of Human Herpesvirus 8-Specific Cytotoxic T-Cell Responses , 1999, Journal of Virology.

[15]  C. Domene,et al.  Structural basis for binding of hypoxia-inducible factor to the oxygen-sensing prolyl hydroxylases. , 2009, Structure.

[16]  H. Ren,et al.  Deletion of immunomodulator C6 from vaccinia virus strain Western Reserve enhances virus immunogenicity and vaccine efficacy , 2013, The Journal of general virology.

[17]  Geoffrey L. Smith,et al.  Steroid Hormone Synthesis by Vaccinia Virus Suppresses the Inflammatory Response to Infection , 2003, The Journal of experimental medicine.

[18]  R. Roepman,et al.  A novel tandem affinity purification strategy for the efficient isolation and characterisation of native protein complexes , 2007, Proteomics.

[19]  H. Ren,et al.  Vaccinia virus protein C4 inhibits NF-κB activation and promotes virus virulence , 2012, The Journal of general virology.

[20]  A. Siddiqui,et al.  Hepatitis C virus stabilizes hypoxia-inducible factor 1alpha and stimulates the synthesis of vascular endothelial growth factor. , 2007, Journal of virology.

[21]  J. Moore,et al.  Steroid hormone synthesis by a vaccinia enzyme: a new type of virus virulence factor. , 1992, The EMBO journal.

[22]  B. Schierwater,et al.  The hypoxia‐inducible transcription factor pathway regulates oxygen sensing in the simplest animal, Trichoplax adhaerens , 2011, EMBO reports.

[23]  A. Le,et al.  Overexpression of Human Papillomavirus Type 16 Oncoproteins Enhances Hypoxia-Inducible Factor 1α Protein Accumulation and Vascular Endothelial Growth Factor Expression in Human Cervical Carcinoma Cells , 2007, Clinical Cancer Research.

[24]  S. Kondo,et al.  EBV latent membrane protein 1 up-regulates hypoxia-inducible factor 1alpha through Siah1-mediated down-regulation of prolyl hydroxylases 1 and 3 in nasopharyngeal epithelial cells. , 2006, Cancer research.

[25]  J. Seong,et al.  Hepatitis B virus X protein enhances transcriptional activity of hypoxia-inducible factor-1alpha through activation of mitogen-activated protein kinase pathway. , 2003, The Journal of biological chemistry.

[26]  P. O’Farrell.  Faculty Opinions recommendation of HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. , 2001 .

[27]  G. Klein,et al.  Epstein-Barr Virus Immortalization of Human B-Cells Leads to Stabilization of Hypoxia-Induced Factor 1 Alpha, Congruent with the Warburg Effect , 2012, PloS one.

[28]  M. Ivan,et al.  HIFα Targeted for VHL-Mediated Destruction by Proline Hydroxylation: Implications for O2 Sensing , 2001, Science.

[29]  Christopher J. Schofield,et al.  Oxygen sensing by HIF hydroxylases , 2004, Nature Reviews Molecular Cell Biology.

[30]  F. Fenner.  Smallpox and its eradication , 1988 .

[31]  K. Khalili,et al.  Oncogenic Viruses and Tumor Glucose Metabolism: Like Kids in a Candy Store , 2012, Molecular Cancer Therapeutics.

[32]  Geoffrey L. Smith,et al.  Vaccinia virus protein C16 acts intracellularly to modulate the host response and promote virulence , 2008, The Journal of general virology.

[33]  B. Moss,et al.  Construction and characterization of an infectious vaccinia virus recombinant that expresses the influenza hemagglutinin gene and induces resistance to influenza virus infection in hamsters. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Heidi L. Kenerson,et al.  Latent Kaposi's Sarcoma-Associated Herpesvirus Infection of Endothelial Cells Activates Hypoxia-Induced Factors , 2006, Journal of Virology.

[35]  M. Karplus,et al.  Evaluation of comparative protein modeling by MODELLER , 1995, Proteins.

[36]  S. Kondo,et al.  Epstein-Barr virus latent membrane protein 1 induces synthesis of hypoxia-inducible factor 1 alpha. , 2004, Molecular and cellular biology.

[37]  M. Sola-Penna,et al.  Herpes simplex type 1 activates glycolysis through engagement of the enzyme 6-phosphofructo-1-kinase (PFK-1). , 2012, Biochimica et biophysica acta.

[38]  G. Semenza,et al.  HIF-1-mediated expression of pyruvate dehydrogenase kinase: a metabolic switch required for cellular adaptation to hypoxia. , 2006, Cell metabolism.

[39]  B. Moss,et al.  Mapping of the vaccinia virus thymidine kinase gene by marker rescue and by cell-free translation of selected mRNA. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[40]  A. Bowie,et al.  Vaccinia Virus Protein C6 Is a Virulence Factor that Binds TBK-1 Adaptor Proteins and Inhibits Activation of IRF3 and IRF7 , 2011, PLoS pathogens.

[41]  B. Moss.  Inhibition of HeLa Cell Protein Synthesis by the Vaccinia Virion , 1968, Journal of virology.

[42]  A. Siddiqui,et al.  Hepatitis C Virus Stabilizes Hypoxia-Inducible Factor 1α and Stimulates the Synthesis of Vascular Endothelial Growth Factor , 2007, Journal of Virology.

[43]  J. Pouysségur,et al.  HIF prolyl‐hydroxylase 2 is the key oxygen sensor setting low steady‐state levels of HIF‐1α in normoxia , 2003, The EMBO journal.

[44]  K. Takeda,et al.  Role and regulation of prolyl hydroxylase domain proteins , 2008, Cell Death and Differentiation.

[45]  Michael I. Wilson,et al.  Targeting of HIF-α to the von Hippel-Lindau Ubiquitylation Complex by O2-Regulated Prolyl Hydroxylation , 2001, Science.