Erythrocytosis-associated HIF-2α Mutations Demonstrate a Critical Role for Residues C-terminal to the Hydroxylacceptor Proline

Furlow, Paul W.; Percy, Melanie J.; Sutherland, Scott; Bierl, Charlene; McMullin, Mary Frances; Master, Stephen R.; Lappin, Terence; Lee, Frank S.

doi:10.1074/jbc.m808737200

Cited by 54 publications

(64 citation statements)

References 33 publications

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“…Families as well as isolated individuals have been encountered in which erythrocytosis owing to elevated plasma Epo levels is explained by heterozygosity for an activating mutation in HIF-2a (Gale et al 2008;Martini et al 2008;Percy et al 2008aPercy et al ,b, 2012Furlow et al 2009;van Wijk et al 2010). These mutations generally impair the ability of HIF-2a to bind to VHL or PHD2.…”

Section: Overproduction Of Epomentioning

confidence: 99%

Erythropoietin

Bunn

2013

Cold Spring Harbor Perspectives in Medicine

208

171

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During the past century, few proteins have matched erythropoietin (Epo) in capturing the imagination of physiologists, molecular biologists, and, more recently, physicians and patients. Its appeal rests on its commanding role as the premier erythroid cytokine, the elegant mechanism underlying the regulation of its gene, and its remarkable impact as a therapeutic agent, arguably the most successful drug spawned by the revolution in recombinant DNA technology. This concise review will begin with a synopsis of the colorful history of this protein, culminating in its purification and molecular cloning. It then covers in more detail the contemporary understanding of Epo's physiology as well as its structure and interaction with its receptor. A major part of this article focuses on the regulation of the Epo gene and the discovery of HIF, a transcription factor that plays a cardinal role in molecular adaptation to hypoxia. In the concluding section, a synopsis of Epo's role in disorders of red blood cell production will be followed by an assessment of the remarkable impact of Epo therapy in the treatment of anemias, as well as concerns that provide a strong impetus for the development of even safer and more effective treatment.

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Section: Overproduction Of Epomentioning

confidence: 99%

Erythropoietin

Bunn

2013

Cold Spring Harbor Perspectives in Medicine

208

171

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“…Some of these mutations cause a reduction in binding to and catalysis by PHD enzymes, as well as a defect in binding to VHL (28). Mutations in PHD2 have also been identified, which cause erythrocytosis in a HIF2␣-dependent manner (27,29).…”

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confidence: 99%

Oxygen-dependent Regulation of Erythropoietin Receptor Turnover and Signaling

Heir

Srikumar

Bikopoulos

et al. 2016

Journal of Biological Chemistry

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von Hippel-Lindau (VHL) disease is a rare familial cancer predisposition syndrome caused by a loss or mutation in a single gene, VHL, but it exhibits a wide phenotypic variability that can be categorized into distinct subtypes. The phenotypic variability has been largely argued to be attributable to the extent of deregulation of the ␣ subunit of hypoxia-inducible factor ␣, a well established target of VHL E3 ubiquitin ligase, ECV (Elongins/ Cul2/VHL). Here, we show that erythropoietin receptor (EPOR) is hydroxylated on proline 419 and 426 via prolyl hydroxylase 3. EPOR hydroxylation is required for binding to the ␤ domain of VHL and polyubiquitylation via ECV, leading to increased EPOR turnover. In addition, several type-specific VHL diseasecausing mutants, including those that have retained proper binding and regulation of hypoxia-inducible factor ␣, showed a severe defect in binding prolyl hydroxylated EPOR peptides. These results identify EPOR as the second bona fide hydroxylation-dependent substrate of VHL that potentially influences oxygen homeostasis and contributes to the complex genotypephenotype correlation in VHL disease.von Hippel-Lindau (VHL) 2 disease is a rare familial cancer predisposition syndrome characterized by the development of tumors in several organ systems, including the central nervous system and retinal hemangioblastoma (HAB), which represent the two cardinal features of VHL disease, as well as endolymphatic sac tumor of the inner ear, pancreatic cystadenoma, benign cystadenoma of epididymis, adnexal papillary cystadenoma of probable mesonephric origin, clear-cell renal cell carcinoma (RCC), paragangliomas, and pheochromocytoma (PHEO) (1). Some VHL patients also develop polycythemia with or without the other stigmata of VHL disease. VHL disease is subcategorized into type 1, characterized by HAB and RCC without PHEO; type 2A, characterized by PHEO and HAB without RCC; type 2B, characterized by PHEO and HAB with RCC; type 2C, characterized by exclusive development of PHEO; and type 3, which is characterized by the development of polycythemia without an increased cancer predisposition (2). Notably, despite the phenotypic heterogeneity, VHL disease is caused by a mutation or loss of a single gene, VHL.VHL is the substrate recognizing subunit of a multiprotein E3 ubiquitin ligase ECV (Elongins BC/Cul2/VHL) that targets the ␣ subunit of hypoxia-inducible factor (HIF␣) for ubiquitylation under normal oxygen tension (3). HIF␣ is hydroxylated on conserved prolines within the oxygen-dependent degradation domain (ODD) by a family of prolyl hydroxylase enzymes (PHD1-3) in the presence of 2-oxoglutarate, iron, and oxygen (4 -7). VHL binds exclusively to prolyl hydroxylated HIF␣. Thus, under hypoxia, the unmodified HIF␣ escapes the recognition by VHL and heterodimerizes with the constitutively expressed HIF␤ to form an active transcription factor, which transactivates numerous hypoxia-inducible genes involved in various adaptive processes to hypoxia such as anaerobic metabolism, angiogenesis, and er...

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“…8 Mutation of Pro531 has been previously shown to result in aberrant stabilization of HIF-2a and thus upregulate genes downstream of HIF transcription. 8 Ala530 is located in the vicinity of the primary hydroxylation site of the HIF-2a. Both Ala530Thr and Ala530Val mutant peptides affect prolyl hydroxylation and VHL protein binding, resulting in reduced HIF-2a degradation but intact transcriptional activity and activation of genes downstream of HIF-2.…”

Section: Discussionmentioning

confidence: 99%

“…PHD hydroxylates specific proline in HIF-a subunits in the context of a strongly conserved LXXLAP sequence motif (where X indicates any amino acid and P indicates the hydroxylacceptor proline). 8 This site-specific proline hydroxylation allows recognition by the VHL protein, a component of an E3 ubiquitin ligase complex that targets hydroxylated HIF-a for degradation by ubiquitin-proteasome pathway. 8 Mutation of Pro531 has been previously shown to result in aberrant stabilization of HIF-2a and thus upregulate genes downstream of HIF transcription.…”

Section: Discussionmentioning

confidence: 99%

“…8 This site-specific proline hydroxylation allows recognition by the VHL protein, a component of an E3 ubiquitin ligase complex that targets hydroxylated HIF-a for degradation by ubiquitin-proteasome pathway. 8 Mutation of Pro531 has been previously shown to result in aberrant stabilization of HIF-2a and thus upregulate genes downstream of HIF transcription. 8 Ala530 is located in the vicinity of the primary hydroxylation site of the HIF-2a.…”

Section: Discussionmentioning

confidence: 99%

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Polycythemia and Paraganglioma With a Novel Somatic HIF2A Mutation in a Male

et al. 2014

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Recently, a new syndrome of paraganglioma, somatostatinoma, and polycythemia has been discovered (known as Pacak-Zhuang syndrome). This new syndrome, with somatic HIF2A gain-of-function mutations, has never been reported in male patients. We describe a male patient with Pacak-Zhuang syndrome who carries a newly discovered HIF2A mutation. Congenital polycythemias have diverse etiologies, including germline mutations in the oxygen-sensing pathway. These include von Hippel-Lindau (Chuvash polycythemia), prolyl hydroxylase domain-containing protein-2, and hypoxia-inducible factor-2a (HIF2a). Somatic gain-of-function mutations in the gene encoding HIF-2a were reported in patients with paraganglioma and polycythemia and have been found exclusively in female patients. Through sequencing of the HIF2A using DNA from paraganglioma in 15-year-old male patient, we identified a novel mutation of HIF2A: a heterozygous C to A substitution at base 1589 in exon 12 of HIF2A. The mutation was not found in germline DNA from leukocytes. The C1589A mutations resulted in substitution of alanine 530 in the HIF-2a protein with glutamic acid. This mutation is undoubtedly associated with increased HIF-2a activity and increased protein half-life, because it affects the vicinity of the prolyl hydroxylase target residue, proline 531. To our knowledge, this is the first report describing Pacak-Zhuang syndrome with somatic gain-of-function mutation in HIF2A in a male patient. Congenital polycythemia of unknown origin should raise suspicion for the novel disorder Pacak-Zhuang syndrome, even in male patients. Pediatrics 2014;133:e1787-e1791

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Erythrocytosis-associated HIF-2α Mutations Demonstrate a Critical Role for Residues C-terminal to the Hydroxylacceptor Proline

Cited by 54 publications

References 33 publications

Erythropoietin

Erythropoietin

Oxygen-dependent Regulation of Erythropoietin Receptor Turnover and Signaling

Polycythemia and Paraganglioma With a Novel Somatic HIF2A Mutation in a Male

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