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The von Hippel-Lindau protein sensitizes renal carcinoma cells to apoptotic stimuli through stabilization of BIMEL

Oncogene volume 28, pages 1864–1874 (2009)Cite this article

Abstract

von Hippel-Lindau (VHL) disease is caused by germ-line mutations in the VHL tumor suppressor gene and is the most common cause of inherited renal cell carcinoma (RCC). Mutations in the VHL gene also occur in a large majority of sporadic cases of clear-cell RCC, which have high intrinsic resistance to chemotherapy and radiotherapy. Here we show that VHL-deficient RCC cells express lower levels of the proapoptotic Bcl-2 family protein BIMEL and are more resistant to etoposide and UV radiation-induced death compared to the same cells stably expressing the wild-type VHL protein (pVHL). Reintroducing pVHL into VHL-null cells increased the half-life of BIMEL protein without affecting its mRNA expression, and overexpressing pVHL inhibited BIMEL polyubiquitination. Suppressing pVHL expression with RNA interference resulted in a decrease in BIMEL protein and a corresponding decrease in the sensitivity of RCC cells to apoptotic stimuli. Directly inhibiting BIMEL expression in pVHL-expressing RCC cells caused a similar decrease in cell death. These results demonstrate that pVHL acts to promote BIMEL protein stability in RCC cells, and that destabilization of BIMEL in the absence of pVHL contributes to the increased resistance of VHL-null RCC cells to certain apoptotic stimuli.

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References

  • Bouillet P, Metcalf D, Huang DC, Tarlinton DM, Kay TW, Kontgen F et al. (1999). Proapoptotic Bcl-2 relative Bim required for certain apoptotic responses, leukocyte homeostasis, and to preclude autoimmunity. Science 286: 1735–1738.

    Article  CAS  Google Scholar 

  • Bruick RK, McKnight SL . (2001). A conserved family of prolyl-4-hydroxylases that modify HIF. Science 294: 1337–1340.

    Article  CAS  Google Scholar 

  • Cohen HT . (1999). Advances in the molecular basis of renal neoplasia. Curr Opin Nephrol Hypertens 8: 325–331.

    Article  CAS  Google Scholar 

  • Devarajan P, De Leon M, Talasazan F, Schoenfeld AR, Davidowitz EJ, Burk RD . (2001). The von Hippel-Lindau gene product inhibits renal cell apoptosis via Bcl-2-dependent pathways. J Biol Chem 276: 40599–40605.

    Article  CAS  Google Scholar 

  • Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O'Rourke J, Mole DR et al. (2001). C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation. Cell 107: 43–54.

    Article  CAS  Google Scholar 

  • Ewings KE, Wiggins CM, Cook SJ . (2007). Bim and the pro-survival Bcl-2 proteins: opposites attract, ERK repels. Cell Cycle 6: 2236–2240.

    Article  CAS  Google Scholar 

  • Frew IJ, Krek W . (2008). pVHL: a multipurpose adaptor protein. Sci Signal 1: pe30.

    Article  Google Scholar 

  • Fu J, Menzies K, Freeman RS, Taubman MB . (2007). EGLN3 prolyl hydroxylase regulates skeletal muscle differentiation and myogenin protein stability. J Biol Chem 282: 12410–12418.

    Article  CAS  Google Scholar 

  • Gnarra JR, Tory K, Weng Y, Schmidt L, Wei MH, Li H et al. (1994). Mutations of the VHL tumour suppressor gene in renal carcinoma. Nat Genet 7: 85–90.

    Article  CAS  Google Scholar 

  • Gorospe M, Egan JM, Zbar B, Lerman M, Geil L, Kuzmin I et al. (1999). Protective function of von Hippel-Lindau protein against impaired protein processing in renal carcinoma cells. Mol Cell Biol 19: 1289–1300.

    Article  CAS  Google Scholar 

  • Hergovich A, Lisztwan J, Barry R, Ballschmieter P, Krek W . (2003). Regulation of microtubule stability by the von Hippel-Lindau tumour suppressor protein pVHL. Nat Cell Biol 5: 64–70.

    Article  CAS  Google Scholar 

  • Iliopoulos O, Kibel A, Gray S, Kaelin Jr WG . (1995). Tumour suppression by the human von Hippel-Lindau gene product. Nat Med 1: 822–826.

    Article  CAS  Google Scholar 

  • Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M et al. (2001). HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 292: 464–468.

    Article  CAS  Google Scholar 

  • Iwai K, Yamanaka K, Kamura T, Minato N, Conaway RC, Conaway JW et al. (1999). Identification of the von Hippel-lindau tumor-suppressor protein as part of an active E3 ubiquitin ligase complex. Proc Natl Acad Sci USA 96: 12436–12441.

    Article  CAS  Google Scholar 

  • Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J, Gaskell SJ et al. (2001). Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. Science 292: 468–472.

    Article  CAS  Google Scholar 

  • Kaelin WG . (2002). Molecular basis of the VHL hereditary cancer syndrome. Nat Rev Cancer 2: 673–682.

    Article  CAS  Google Scholar 

  • Kim M, Yan Y, Lee K, Sgagias M, Cowan KH . (2004). Ectopic expression of von Hippel-Lindau tumor suppressor induces apoptosis in 786-O renal cell carcinoma cells and regresses tumor growth of 786-O cells in nude mouse. Biochem Biophys Res Commun 320: 945–950.

    Article  CAS  Google Scholar 

  • Kurban G, Duplan E, Ramlal N, Hudon V, Sado Y, Ninomiya Y et al. (2008). Collagen matrix assembly is driven by the interaction of von Hippel-Lindau tumor suppressor protein with hydroxylated collagen IV alpha 2. Oncogene 27: 1004–1012.

    Article  CAS  Google Scholar 

  • Lee S, Nakamura E, Yang H, Wei W, Linggi MS, Sajan MP et al. (2005). Neuronal apoptosis linked to EglN3 prolyl hydroxylase and familial pheochromocytoma genes: developmental culling and cancer. Cancer Cell 8: 155–167.

    Article  Google Scholar 

  • Ley R, Balmanno K, Hadfield K, Weston C, Cook SJ . (2003). Activation of the ERK1/2 signaling pathway promotes phosphorylation and proteasome-dependent degradation of the BH3-only protein, Bim. J Biol Chem 278: 18811–18816.

    Article  CAS  Google Scholar 

  • Ley R, Ewings KE, Hadfield K, Cook SJ . (2005). Regulatory phosphorylation of Bim: sorting out the ERK from the JNK. Cell Death Differ 12: 1008–1014.

    Article  CAS  Google Scholar 

  • Lipscomb EA, Sarmiere PD, Crowder RJ, Freeman RS . (1999). Expression of the SM-20 gene promotes death in nerve growth factor-dependent sympathetic neurons. J Neurochem 73: 429–432.

    Article  CAS  Google Scholar 

  • Lonergan KM, Iliopoulos O, Ohh M, Kamura T, Conaway RC, Conaway JW et al. (1998). Regulation of hypoxia-inducible mRNAs by the von Hippel-Lindau tumor suppressor protein requires binding to complexes containing elongins B/C and Cul2. Mol Cell Biol 18: 732–741.

    Article  CAS  Google Scholar 

  • Maxwell PH, Wiesener MS, Chang GW, Clifford SC, Vaux EC, Cockman ME et al. (1999). The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis. Nature 399: 271–275.

    Article  CAS  Google Scholar 

  • Nyhan MJ, O'Sullivan GC, McKenna SL . (2008). Role of the VHL (von Hippel-Lindau) gene in renal cancer: a multifunctional tumour suppressor. Biochem Soc Trans 36: 472–478.

    Article  CAS  Google Scholar 

  • O'Connor L, Strasser A, O'Reilly LA, Hausmann G, Adams JM, Cory S et al. (1998). Bim: a novel member of the Bcl-2 family that promotes apoptosis. EMBO J 17: 384–395.

    Article  CAS  Google Scholar 

  • Okuda H, Saitoh K, Hirai S, Iwai K, Takaki Y, Baba M et al. (2001). The von Hippel-Lindau tumor suppressor protein mediates ubiquitination of activated atypical protein kinase C. J Biol Chem 276: 43611–43617.

    Article  CAS  Google Scholar 

  • Qi H, Ohh M . (2003). The von Hippel-Lindau tumor suppressor protein sensitizes renal cell carcinoma cells to tumor necrosis factor-induced cytotoxicity by suppressing the nuclear factor-kappaB-dependent antiapoptotic pathway. Cancer Res 63: 7076–7080.

    CAS  PubMed  Google Scholar 

  • Qi XJ, Wildey GM, Howe PH . (2006). Evidence that Ser87 of BimEL is phosphorylated by Akt and regulates BimEL apoptotic function. J Biol Chem 281: 813–823.

    Article  CAS  Google Scholar 

  • Roe JS, Kim H, Lee SM, Kim ST, Cho EJ, Youn HD . (2006). p53 stabilization and transactivation by a von Hippel-Lindau protein. Mol Cell 22: 395–405.

    Article  CAS  Google Scholar 

  • Schermer B, Ghenoiu C, Bartram M, Muller RU, Kotsis F, Hohne M et al. (2006). The von Hippel-Lindau tumor suppressor protein controls ciliogenesis by orienting microtubule growth. J Cell Biol 175: 547–554.

    Article  CAS  Google Scholar 

  • Schoenfeld AR, Parris T, Eisenberger A, Davidowitz EJ, De Leon M, Talasazan F et al. (2000). The von Hippel-Lindau tumor suppressor gene protects cells from UV-mediated apoptosis. Oncogene 19: 5851–5857.

    Article  CAS  Google Scholar 

  • Semenza GL . (2003). Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3: 721–732.

    Article  CAS  Google Scholar 

  • Stebbins CE, Kaelin Jr WG, Pavletich NP . (1999). Structure of the VHL-ElonginC-ElonginB complex: implications for VHL tumor suppressor function. Science 284: 455–461.

    Article  CAS  Google Scholar 

  • van Houwelingen KP, van Dijk BA, Hulsbergen-van de Kaa CA, Schouten LJ, Gorissen HJ, Schalken JA et al. (2005). Prevalence of von Hippel-Lindau gene mutations in sporadic renal cell carcinoma: results from The Netherlands cohort study. BMC Cancer 5: 57.

    Article  Google Scholar 

  • Willis SN, Fletcher JI, Kaufmann T, van Delft MF, Chen L, Czabotar PE et al. (2007). Apoptosis initiated when BH3 ligands engage multiple Bcl-2 homologs, not Bax or Bak. Science 315: 856–859.

    Article  CAS  Google Scholar 

  • Xie L, Johnson RS, Freeman RS . (2005). Inhibition of NGF deprivation-induced death by low oxygen involves suppression of BIM(EL) and activation of HIF-1. J Cell Biol 168: 911–920.

    Article  CAS  Google Scholar 

  • Yagoda A, Abi-Rached B, Petrylak D . (1995). Chemotherapy for advanced renal-cell carcinoma: 1983-1993. Semin Oncol 22: 42–60.

    CAS  PubMed  Google Scholar 

  • Yang H, Minamishima YA, Yan Q, Schlisio S, Ebert BL, Zhang X et al. (2007). pVHL acts as an adaptor to promote the inhibitory phosphorylation of the NF-kappaB agonist Card9 by CK2. Mol Cell 28: 15–27.

    Article  Google Scholar 

  • Zantl N, Weirich G, Zall H, Seiffert BM, Fischer SF, Kirschnek S et al. (2007). Frequent loss of expression of the pro-apoptotic protein Bim in renal cell carcinoma: evidence for contribution to apoptosis resistance. Oncogene 26: 7038–7048.

    Article  CAS  Google Scholar 

  • Zhang W, Cheng GZ, Gong J, Hermanto U, Zong CS, Chan J et al. (2008). RACK1 and CIS Mediate the Degradation of BimEL in Cancer Cells. J Biol Chem 283: 16416–16426.

    Article  CAS  Google Scholar 

  • Zhou MI, Wang H, Ross JJ, Kuzmin I, Xu C, Cohen HT . (2002). The von Hippel-Lindau tumor suppressor stabilizes novel plant homeodomain protein Jade-1. J Biol Chem 277: 39887–39898.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr William Kaelin for providing cell lines and Dr Thomas Benzing and Dr Patricia Hinkle for providing plasmids used in this study. This work was supported by grants NS034400 and NS058868 from the National Institutes of Health.

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Author notes

  1. M C Schoell

    Present address: 3Current address: Department of Anatomy and Neurobiology, Neuroscience Institute, Morehouse School of Medicine, Atlanta, GA, USA.,

Authors and Affiliations

  1. Interdepartmental Graduate Program in Neuroscience, University of Rochester School of Medicine, Rochester, NY, USA

    Y Guo

  2. Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, NY, USA

    M C Schoell & R S Freeman

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Correspondence to R S Freeman.

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Guo, Y., Schoell, M. & Freeman, R. The von Hippel-Lindau protein sensitizes renal carcinoma cells to apoptotic stimuli through stabilization of BIMEL. Oncogene 28, 1864–1874 (2009). https://doi.org/10.1038/onc.2009.35

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