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Published ahead of print on June 14, 2006
J Am Soc Nephrol 17: 1801-1806, 2006
© 2006 American Society of Nephrology
doi: 10.1681/ASN.2006020181
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Formation of Primary Cilia in the Renal Epithelium Is Regulated by the von Hippel-Lindau Tumor Suppressor Protein

Miguel A. Esteban, Sarah K. Harten, Maxine G. Tran and Patrick H. Maxwell

Renal Laboratory, Imperial College London, Hammersmith Campus, London, United Kingdom

Address correspondence to: Dr. Patrick H. Maxwell, Renal Laboratory, Imperial College London, Hammersmith Campus, Du Cane Road, London W12 0NN, UK. Phone: +44-20-8383-8594; Fax: +44-20-8383-2062; p.maxwell{at}imperial.ac.uk


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results and Discussion
 Conclusion
 References
 
Growing evidence points to defects in the primary cilium as a critical mechanism underlying renal cyst development. Inactivation of the VHL gene is responsible for the autosomal dominant condition von Hippel-Lindau (VHL) disease and is implicated in most sporadic clear cell renal carcinomas. Manifestations of VHL disease include cysts in several organs, particularly in the kidney. Here it is shown that VHL inactivation is associated with abrogation of the primary cilium in renal cysts of patients with VHL disease and in VHL-defective cell lines. Complementation of VHL-defective clear cell renal carcinoma cell lines with wild-type VHL restored primary cilia. Moreover, it is shown that the effects of VHL on the primary cilium are mediated substantially via hypoxia-inducible factor. The effect of VHL status on the primary cilium provides a potential mechanism for renal cyst development in VHL disease and may help in the understanding of how VHL acts as a tumor suppressor.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results and Discussion
 Conclusion
 References
 
Many different hereditary conditions are associated with development of renal cysts, often with other clinical manifestations. These include autosomal dominant polycystic kidney disease, Bardet-Biedl syndrome, nephronophthisis, and oral-facial-digital type 1 syndrome. Remarkably, a common link has emerged in that mutations in the genes underlying these cystic conditions alter the structure or function of the primary cilium (1–7), a luminal hair-like extracellular appendage that transmits calcium-mediated intracellular signals after mechanical bending (8,9). These calcium signals are thought to regulate cytoarchitecture and cellular proliferation of renal tubular cells in response to urine flow.

von Hippel-Lindau (VHL) disease is an uncommon autosomal dominant condition that is caused by inheritance of a mutant VHL allele; the main renal manifestations are a very high risk for developing clear cell renal carcinomas (CCRCC) and renal cysts. Other manifestations include pheochromocytoma; hemangioblastomas in the retina, cerebellum, and spinal cord; endolymphatic sac tumors; and epididymal cysts. The VHL gene is situated at 3p25, and a large number of different mutations have been identified in kindreds with VHL disease. VHL behaves as a classic two-hit tumor suppressor gene that conforms to Knudson’s model (10), with the clinical manifestations (including renal cysts and tumors) invariably involving somatic inactivation of the remaining wild-type VHL allele. VHL also is inactivated in the majority of sporadic CCRCC, which is the most common type of renal cancer. Importantly, re-expression of VHL in cell lines that are derived from CCRCC suppresses their tumorigenicity in nude mice (11). In view of the proposed role of the primary cilium in other kidney cystic diseases, we hypothesized that the VHL protein (pVHL) may influence the formation, maintenance, and/or function of the primary cilium.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results and Discussion
 Conclusion
 References
 
Imaging Techniques
Immunohistochemistry and immunofluorescence microscopy were performed as described previously (12,13). For immunofluorescence, we also used a laser scanning confocal microscope (Zeiss LSM 5 PASCAL, Carl Zeiss, Oberkochen, Germany) equipped with Zeiss LSM image browser version 3.2.0.115. Scanning electron microscopy was performed using a Jeol microscope (Akishima, Japan) at University College London, courtesy of Prof. K. Matter.

Cells and Antibodies
RCC4, RCC10, and sublines were described previously (13,14). For imaging experiments, cells were plated to confluence and studied after 3 or 4 d.

Anti–hypoxia-inducible factor 1{alpha} (anti–HIF-1{alpha}) was purchased from Transduction Labs (Lexington, KY), anti–HIF-2{alpha} was purchased from Cancer Research UK (London, UK), anti–{alpha}-tubulin and antiacetylated {alpha}-tubulin were purchased from Sigma (St. Louis, MO), and anti–carbonic anhydrase IX (anti-CAIX) was a gift from S. Pastorekova (Institute of Virology, Bratislava, Slovak Republic).

Retroviral Infection and Plasmids
pCMVR-VHL N78S was constructed by transferring an insert from a pcDNA3 plasmid (gift of W. Krek [Institute of Cell Biology, Zurich, Switzerland] and A. Hergovich [Friedrich Miescher Institute, Basel, Switzerland]). Other retroviral vectors and procedures for infection were described previously (13,15).

Quantitative Real-Time Reverse Transcription–PCR
PCR analysis using SYBR Green (AB gene) was performed as described previously (13). All real-time reverse transcription–PCR data are given as a value normalized to the level of beta-actin expression in the same retrotranscription. Sequences for primers are available on request.

siRNA Transfection
Transfection procedures and oligo sequences were described previously (13,16).


   Results and Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
Conclusion
 References
 
To study whether VHL status may influence the primary cilium, we first performed immunohistochemistry for acetylated {alpha}-tubulin (an essential constituent of the primary cilium) on paraffin-embedded material that contained cysts from the kidneys of two different patients with VHL disease. In the same section, normal kidney tubules showed primary cilia protruding from the luminal side of epithelial cells (Figure 1, left), but these structures could not be identified in cysts (Figure 1, right). Adjacent sections of the cyst wall showed strong positive labeling for CAIX (Figure 1, right), consistent with biallelic inactivation of VHL in the cells that line the cyst (12).


Figure 1
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  		Figure 1. Primary cilia are lost in renal cysts from patients with von Hippel-Lindau (VHL) disease. Serial sections from a single block that contained a cyst and normal renal tubules were labeled for acetylated {alpha}-tubulin (top) and carbonic anhydrase IX (CAIX; a marker of biallelic inactivation of VHL; bottom). Middle panels show sections with low magnification; left and right panels show the indicated areas in the central panels at increased magnification. In the normal tubules (left), primary cilia (indicated with arrowheads) are visible, and CAIX is not expressed. In the cyst (right), primary cilia are not seen, and the cells express CAIX.

 
Next, we used two different pVHL-negative CCRCC cell lines (RCC4 and RCC10) and corresponding isogenic sublines that express pVHL (hereafter referred to as RCC4/VHL and RCC10/VHL). Cells were studied 3 to 4 d after reaching confluence, which also is necessary for the development of cilia in MDCK cells (7). Remarkably, immunofluorescence microscopy for acetylated {alpha}-tubulin readily detected abundant primary cilia only in the CCRCC cells that expressed pVHL (Figure 2A). In all four cell lines, labeling for acetylated {alpha}-tubulin also was present in the cytoplasm, with pVHL-negative cells showing somewhat more cytoplasmic signal. When pVHL-negative RCC4 and RCC10 cultures were studied at longer intervals after reaching confluence, some rudimentary cilia were visible, but these were always sparse and much less well developed than those seen in RCC4/VHL and RCC10/VHL. The presence of primary cilia in RCC4/VHL and RCC10/VHL also was verified using confocal microscopy (Figure 2B). Similar results were obtained using scanning electron microscopy (Figure 2C). Consistent with the immunofluorescence, pVHL-defective RCC4 and RCC10 cells displayed very few or no cilia on confocal microscopy (Figure 2B) and scanning electron microscopy (data not shown).


Figure 2
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  		Figure 2. Re-expression of VHL protein (pVHL) in clear cell renal carcinoma (CCRCC) cells restores cilia. (A) Immunofluorescence for acetylated {alpha}-tubulin in RCC4 and RCC10 cells. Arrow in the bottom panel indicates primary cilia in RCC4/VHL cells. Bars = 20 µm. (B) Vertical computer reconstruction (using a confocal immunofluorescence microscope) of RCC4 and RCC10 cells and stable transfectants that expressed pVHL. *Primary cilia. Bars = 20 µm. (C) Scanning electron microscopy of CCRCC cells complemented with pVHL. Bars = 5 and 10 µm.

 
pVHL has been reported to have a number of biochemical functions, including regulation of microtubule stability, cell differentiation, cell motility, extracellular matrix assembly, JunB, and atypical isoforms of protein kinase C (15,17–22). However, the best characterized function of pVHL is to act as an essential component in the degradation of HIF-{alpha} subunits (10,14). In the presence of oxygen, pVHL captures {alpha} subunits of the transcription factor HIF, resulting in their degradation by the proteasome. The molecular signal for pVHL-mediated capture is the hydroxylation of two prolyl residues in the central part of HIF-{alpha} subunits, by a family of oxygen-dependent dioxygenases, PHD1-3 (prolyl hydroxylase domain–containing proteins) (23). When oxygenation is reduced or pVHL is absent, HIF becomes stabilized and promotes the transcription of multiple target genes that are involved in diverse pathways, examples of which are erythropoiesis, angiogenesis, glucose uptake, and glycolysis (24). Broadly, these actions can be seen as adapting the cell, tissue, or whole organism to low oxygen. There are three different HIF-{alpha} isoforms (HIF-1{alpha}, HIF-2{alpha}, and HIF-3{alpha}), the best characterized of which are HIF-1{alpha} and HIF-2{alpha} (10). HIF-1{alpha} and HIF-2{alpha} are not redundant, based on genetic inactivation experiments in mice. However, their relative roles in responses to hypoxia are not yet completely understood. It is interesting that in CCRCC cell lines, HIF-2{alpha} is necessary for tumorigenesis in xenograft models and selectively increases Cyclin D1 expression (25,26), whereas HIF-1{alpha} selectively increases expression of CAIX and the proapoptotic gene BNIP3 (25).

We hypothesized that constitutive activation of HIF may be the mechanism underlying altered ciliogenesis when VHL is inactivated. To dissect this, we first used retroviral gene transfer to express several different pVHL molecules. These included the full-length p30 isoform (pVHL30) and the p19 isoform (pVHL19). The latter isoform also is able to regulate HIF and arises from an alternative translation initiation site at amino acid 54 (10). We also tested two disease-associated missense mutations in pVHL, resulting in single amino acid substitutions, pVHL-V84L and pVHL-L188V. These mutations are associated with type 2C VHL disease, in which patients develop pheochromocytoma without other clinical manifestations (10). pVHL type 2C mutations retain the ability to regulate HIF normally (27,28), thereby providing a useful tool to identify specific consequences of pVHL loss of function that are independent of HIF (29,30). As expected, expression of pVHL19, pVHL30, or either of the two pVHL type 2C mutants (but not the empty vector) comparably reduced protein levels of HIF-{alpha} subunits in infected pools of RCC4 and RCC10 cells (Figure 3A); quantitative real-time reverse transcription–PCR analysis also showed comparable reduction in glucose transporter 1 (a well-characterized HIF target) mRNA levels (Figure 3B). Notably, immunofluorescence for acetylated {alpha}-tubulin showed that formation of the primary cilium was restored in RCC4 and RCC10 cells that were infected with pVHL19, pVHL30, and also the two pVHL 2C mutations (Figure 3C). Infection of pVHL-negative cells with a mutant VHL gene encoding a different missense substitution that abolishes the ability to regulate HIF (VHL N78S) (27) was used as an additional control and did not result in formation of cilia (Figure 3D). These experiments show that restoration of the primary cilium shown in stably transfected RCC4/VHL and RCC10/VHL cells (see Figure 2A) is not due to clone-specific effects, because it also was seen in these freshly prepared heterogeneous pooled populations.


Figure 3
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  		Figure 3. Activation of hypoxia-inducible factor {alpha} (HIF-1{alpha}) underlies the abrogation of primary cilia in CCRCC cells. (A) Western blotting shows comparable suppression of HIF-1{alpha} and HIF-2{alpha} in cells infected with wild-type pVHL or the type 2C mutations. Expression of wild-type and mutant pVHL proteins was confirmed by Western blot analysis (data not shown) (13). (B) Quantitative real-time reverse transcription–PCR (RT-PCR) of glucose transporter 1 (GLUT1) mRNA. (C) Immunofluorescence for acetylated {alpha}-tubulin. Bars = 20 µm. (D) Western blotting (HIF-1{alpha}, HIF-2{alpha}, and {alpha}-tubulin), quantitative real-time RT-PCR of GLUT1 mRNA, and immunofluorescence for acetylated {alpha}-tubulin (bars = 20 µm) in RCC4 cells infected with empty vector, pVHL30, or pVHL N78S. (E) siRNA treatment of pVHL-negative RCC10 cells with specific oligos directed against HIF-1{alpha}, HIF-2{alpha}, or firefly luciferase (control). Western blotting shows specific inhibition of HIF-1{alpha} and HIF-2{alpha}. Immunofluorescence microscopy for acetylated {alpha}-tubulin shows restoration of primary cilia on knocking down HIF-1{alpha}. Bars = 20 µm. (F) pVHL stably transfected RCC10 cells infected with retrovirus that encodes a constitutively active form of HIF-1{alpha} or empty vector. Western blotting verified the corresponding presence of HIF-1{alpha}. Immunofluorescence microscopy for acetylated {alpha}-tubulin in the same cells. Bars = 20 µm.

 
The concordance between the ability of pVHL molecules to suppress HIF and restore the primary cilium supported the notion that the mechanism involved activation of HIF. To test this directly, we then performed genetic knockdown of HIF-1{alpha} or HIF-2{alpha} using siRNA treatment of pVHL-negative RCC10 cells. Specificity was confirmed by Western blotting (Figure 3D); siRNA knockdown of HIF-2{alpha} resulted in a reproducible increase in HIF-1{alpha} as reported previously (13,25). It is interesting that immunofluorescence microscopy for acetylated {alpha}-tubulin showed that only knockdown of HIF-1{alpha} significantly restored cilium formation (Figure 3E). To demonstrate further the role of HIF-1{alpha} in preventing renal ciliogenesis, we also infected RCC10/VHL with retroviruses that encode constitutively active forms of HIF-1{alpha}; infection with the empty vector was used as a control. The corresponding presence of HIF-1{alpha} was verified by Western blotting (Figure 3F). Notably, immunofluorescence staining showed that compared with control cells, active HIF-1{alpha} altered the formation of the cilia (Figure 3F). Therefore, suppression of HIF-1{alpha} restores cilium formation in pVHL-negative CCRCC cells, and activation of HIF-1{alpha} in CCRCC cells that re-express pVHL abolishes it.

Understanding how pVHL regulates the primary cilium via HIF-1{alpha} will require further work, but it is intriguing to speculate how this may occur. It is possible that the effect on cilia is because pVHL is required for normal interactions between adjacent cells and/or with the extracellular matrix, both of which are important in differentiation decisions (14–18). In this regard, we recently reported that in pVHL-negative CCRCC cells, HIF regulates expression of E-cadherin (13), a cell–cell adhesion molecule that is responsible for controlling differentiation in multiple cell models. It will be interesting to determine whether forced expression of E-cadherin can restore cilium formation in pVHL-negative CCRCC cells. Another interesting possibility is that HIF-1{alpha} could have a direct effect on the expression of a constituent of the cilium or a gene that is required for its formation and function. Maintenance of the primary cilium structure is mediated by a bidirectional cargo process termed intraflagellar transport (31), which is highly energy consuming. It therefore could be envisaged that activation of HIF-1{alpha} in hypoxia (and pVHL-negative cells) would shut down intraflagellar transport as a strategy to conserve energy for other cellular functions.


   Conclusion
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 Conclusion
References
 
Our study establishes a link between the formation of renal cysts in VHL disease and the pathogenesis of other hereditary kidney cystic diseases, thereby providing further support for the "ciliary hypothesis" (31). Potentially, progress toward understanding other cystic kidney diseases will shed light on how cysts develop in VHL disease and vice versa. Our work also provides a link between activation of the HIF transcription factor and disturbance of the cilium. This is important because activation of HIF theoretically could provide a route by which reduced oxygen levels in other pathologies in the kidney would induce cyst formation; for example, it may contribute to cyst formation in patients with longstanding renal disease.


   Acknowledgments
 
This work was funded by the Wellcome Trust, Cancer Research UK, the Medical Research Council, and the EU framework 6 integrated project Euroxy. Miguel A. Esteban was funded by a Traveling Research Fellowship from the Wellcome Trust.


   Footnotes
 
Published online ahead of print. Publication date available at www.jasn.org.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 Conclusion
 References
 

  1. Kulaga HM, Leitch CC, Eichers ER, Badano JL, Lesemann A, Hoskins BE, Lupski JR, Beales PL, Reed RR, Katsanis N: Loss of BBS proteins causes anosmia in humans and defects in olfactory cilia structure and function in the mouse. Nat Genet 9: 994–998, 2004
  2. Otto EA, Loeys B, Khanna H, Hellemans J, Sudbrak R, Fan S, Muerb U, O’Toole JF, Helou J, Attanasio M, Utsch B, Sayer JA, Lillo C, Jimeno D, Coucke P, De Paepe A, Reinhardt R, Klages S, Tsuda M, Kawakami I, Kusakabe T, Omran H, Imm A, Tippens M, Raymond PA, Hill J, Beales P, He S, Kispert A, Margolis B, Williams DS, Swaroop A, Hildebrandt F: Nephrocystin-5, a ciliary IQ domain protein, is mutated in Senior-Loken syndrome and interacts with RPGR and calmodulin. Nat Genet 37: 282–288, 2005[CrossRef][Medline]
  3. Ansley SJ, Badano JL, Blacque OE, Hill J, Hoskins BE, Leitch CC, Kim JC, Ross AJ, Eichers ER, Teslovich TM, Mah AK, Johnsen RC, Cavender JC, Lewis RA, Leroux MR, Beales PL, Katsanis N: Basal body dysfunction is a likely cause of pleiotropic Bardet-Biedl syndrome. Nature 425: 628–633, 2003[CrossRef][Medline]
  4. Ferrante MI, Zullo A, Barra A, Bimonte S, Messaddeq N, Studer M, Dolle P, Franco B: Oral-facial-digital type I protein is required for primary cilia formation and left-right axis specification. Nat Genet 38: 112–117, 2006[Medline]
  5. Davenport JR, Yoder BK: An incredible decade for the primary cilium: A look at a once-forgotten organelle. Am J Physiol Renal Physiol 289: F1159–F1169, 2005[Abstract/Free Full Text]
  6. Pazour GJ, Dickert BL, Vucica Y, Seeley ES, Rosenbaum JL, Witman GB, Cole DG: Chlamydomonas IFT88 and its mouse homologue, polycystic kidney disease gene tg737, are required for assembly of cilia and flagella. J Cell Biol 151: 709–718, 2000[Abstract/Free Full Text]
  7. Murcia NS, Richards WG, Yoder BK, Mucenski ML, Dunlap JR, Woychik RP: The Oak Ridge Polycystic Kidney (orpk) disease gene is required for left-right axis determination. Development 127: 2347–2355, 2000[Abstract]
  8. Nauli SM, Alenghat FJ, Luo Y, Williams E, Vassilev P, Li X, Elia AE, Lu W, Brown EM, Quinn SJ, Ingber DE, Zhou J: Polycystins 1 and 2 mediate mechanosensation in the primary cilium of kidney cells. Nat Genet 33: 129–137, 2003[CrossRef][Medline]
  9. Praetorius HA, Spring KR: Removal of the MDCK cell primary cilium abolishes flow sensing. J Membr Biol 191: 69–76, 2003[CrossRef][Medline]
  10. Kaelin WG Jr: Molecular basis of the VHL hereditary cancer syndrome. Nat Rev Cancer 2: 673–682, 2002[CrossRef][Medline]
  11. Iliopoulos O, Kibel A, Gray S, Kaelin WG Jr: Tumour suppression by the human von Hippel-Lindau gene product. Nat Med 1: 822–826, 1995[CrossRef][Medline]
  12. Mandriota SJ, Turner KJ, Davies DR, Murray PG, Morgan NV, Sowter HM, Wykoff CC, Maher ER, Harris AL, Ratcliffe PJ, Maxwell PH: HIF activation identifies early lesions in VHL kidneys: Evidence for site-specific tumor suppressor function in the nephron. Cancer Cell 1: 459–468, 2002[CrossRef][Medline]
  13. Esteban MA, Tran MG, Harten S, Hill P, Castellanos MC, Chandra A, Raval R, O’Brien TS, Maxwell PH: Regulation of E-cadherin expression by VHL and hypoxia-inducible factor. Cancer Res 66: 3567–3575, 2006[Abstract/Free Full Text]
  14. Maxwell PH, Wiesener MS, Chang GW, Clifford SC, Vaux EC, Cockman ME, Wykoff CC, Pugh CW, Maher ER, Ratcliffe PJ: The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis. Nature 399: 271–275, 1999[CrossRef][Medline]
  15. Hergovich A, Lisztwan J, Barry R, Ballschmieter P, Krek W: Regulation of microtubule stability by the von Hippel-Lindau tumour suppressor protein pVHL. Nat Cell Biol 5: 64–70, 2003[CrossRef][Medline]
  16. Sowter HM, Raval RR, Moore JW, Ratcliffe PJ, Harris AL: Predominant role of hypoxia-inducible transcription factor (Hif)-1alpha versus Hif-2alpha in regulation of the transcriptional response to hypoxia. Cancer Res 63: 6130–6134, 2003[Abstract/Free Full Text]
  17. Ohh M, Yauch RL, Lonergan KM, Whaley JM, Stemmer-Rachamimov AO, Louis DN, Gavin BJ, Kley N, Kaelin WG Jr, Iliopoulos O: The von Hippel-Lindau tumor suppressor protein is required for proper assembly of an extracellular fibronectin matrix. Mol Cell 1: 959–968, 1998[CrossRef][Medline]
  18. Davidowitz EJ, Schoenfeld AR, Burk RD: VHL induces renal cell differentiation and growth arrest through integration of cell-cell and cell-extracellular matrix signaling. Mol Cell Biol 21: 865–874, 2001[Abstract/Free Full Text]
  19. Baba M, Hirai S, Kawakami S, Kishida T, Sakai N, Kaneko S, Yao M, Shuin T, Kubota Y, Hosaka M, Ohno S: Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth. Oncogene 20: 2727–2736, 2001[CrossRef][Medline]
  20. Esteban-Barragan MA, Avila P, Alvarez-Tejado M, Gutierrez MD, Garcia-Pardo A, Sanchez-Madrid F, Landazuri MO: Role of the von Hippel-Lindau tumor suppressor gene in the formation of beta1-integrin fibrillar adhesions. Cancer Res 62: 2929–2936, 2002[Abstract/Free Full Text]
  21. Kamada M, Suzuki K, Kato Y, Okuda H, Shuin T: von Hippel-Lindau protein promotes the assembly of actin and vinculin and inhibits cell motility. Cancer Res 61: 4184–4189, 2001[Abstract/Free Full Text]
  22. Okuda H, Saitoh K, Hirai S, Iwai K, Takaki Y, Baba M, Minato N, Ohno S, Shuin T: The von Hippel-Lindau tumor suppressor protein mediates ubiquitination of activated atypical protein kinase C. J Biol Chem 276: 43611–43617, 2001[Abstract/Free Full Text]
  23. Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O’Rourke J, Mole DR, Mukherji M, Metzen E, Wilson MI, Dhanda A, Tian YM, Masson N, Hamilton DL, Jaakkola P, Barstead R, Hodgkin J, Maxwell PH, Pugh CW, Schofield CJ, Ratcliffe PJ: C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation. Cell 107: 43–54, 2001[CrossRef][Medline]
  24. Semenza GL: Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3: 721–732, 2003[CrossRef][Medline]
  25. Raval RR, Lau KW, Tran MG, Sowter HM, Mandriota SJ, Li JL, Pugh CW, Maxwell PH, Harris AL, Ratcliffe PJ: Contrasting properties of hypoxia-inducible factor 1 (HIF-1) and HIF-2 in von Hippel-Lindau-associated renal cell carcinoma. Mol Cell Biol 25: 5675–6686, 2005[Abstract/Free Full Text]
  26. Kondo K, Klco J, Nakamura E, Lechpammer M, Kaelin WG Jr: Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein. Cancer Cell 1: 37–46, 2002[CrossRef][Medline]
  27. Clifford SC, Cockman ME, Smallwood AC, Mole DR, Woodward ER, Maxwell PH, Ratcliffe PJ, Maher ER: Contrasting effects on HIF-1alpha regulation by disease-causing pVHL mutations correlate with patterns of tumourigenesis in von Hippel-Lindau disease. Hum Mol Genet 10: 1029–1038, 2001[Abstract/Free Full Text]
  28. Hoffman MA, Ohh M, Yang H, Klco JM, Ivan M, Kaelin WG Jr: von Hippel-Lindau protein mutants linked to type 2C VHL disease preserve the ability to downregulate HIF. Hum Mol Genet 10: 1019–1027, 2001[Abstract/Free Full Text]
  29. Stickle NH, Chung J, Klco JM, Hill RP, Kaelin WG Jr, Ohh M: pVHL modification by NEDD8 is required for fibronectin matrix assembly and suppression of tumor development. Mol Cell Biol 24: 3251–3261, 2004[Abstract/Free Full Text]
  30. Lee S, Nakamura E, Yang H, Wei W, Linggi MS, Sajan MP, Farese RV, Freeman RS, Carter BD, Kaelin WG Jr, Schlisio S: Neuronal apoptosis linked to EglN3 prolyl hydroxylase and familial pheochromocytoma genes: Developmental culling and cancer. Cancer Cell 8: 155–167, 2005[CrossRef][Medline]
  31. Pazour GJ: Intraflagellar transport and cilia-dependent renal disease: The ciliary hypothesis of polycystic kidney disease. J Am Soc Nephrol 15: 2528–2536, 2004[Abstract/Free Full Text]



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