HIF-1α-prolyl hydroxylase: molecular target of nitric oxide in the hypoxic signal transduction pathway

Wang, Feng; Sekine, Hiroshi; Kikuchi, Yasuo; Takasaki, Chikahisa; Miura, Chisa; Okuda, Heiwa; Shuin, Taro; Fujii‐Kuriyama, Yoshiaki; Sogawa, Kazuhiro

doi:10.1016/s0006-291x(02)00729-5

Cited by 61 publications

(33 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several other agents have been reported to lower HIF-1a protein levels, including topoisomerase I and II inhibitors (38, 39), phosphatidylinositol 3-kinase inhibitors (40), heat shock protein 90 inhibitors (41), microtubule cytoskeleton disrupting agents (42), HIV protease inhibitors (43), and YC-1, an activator of soluble guanyl cyclase that stimulates prolyl hydroxylase activity (44). None of these agents is selective for HIF-1a and the contribution of HIF-1a inhibition to their activity has not been established.…”

Section: Discussionmentioning

confidence: 99%

Molecular mechanisms for the activity of PX-478, an antitumor inhibitor of the hypoxia-inducible factor-1α

Koh

Spivak-Kroizman

Venturini

et al. 2008

Molecular Cancer Therapeutics

266

188

View full text Add to dashboard Cite

We have reported previously that PX-478 (S-2-amino-3-[4 ¶-N,N,-bis(chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride) has potent antitumor activity against a variety of human tumor xenografts associated with the levels of the hypoxia-inducible factor-1A (HIF-1A) within the tumor. We now report that PX-478 inhibits HIF-1A protein levels and transactivation in a variety of cancer cell lines. Hypoxia-induced vascular endothelial growth factor formation was inhibited by PX-478, whereas baseline levels of vascular endothelial growth factor in normoxia were unaffected. Studies of the mechanism of PX-478 action showed that HIF-1A inhibition occurs in both normoxia and hypoxia and does not require pVHL or p53. In addition, PX-478 decreases levels of HIF-1A mRNA and inhibits translation as determined by 35 S labeling experiments and reporter assays using the 5 ¶ untranslated region of HIF-1A. Moreover, to a lesser extent, PX-478 also inhibits HIF-1A deubiquitination resulting in increased levels of polyubiquitinated HIF-1A. The inhibitory effect of PX-478 on HIF-1A levels is primarily due to its inhibition of translation because HIF-1A translation continues in hypoxia when translation of most proteins is decreased. We conclude that PX-478 inhibits HIF-1A at multiple levels that together or individually may contribute to its antitumor activity against HIF-1A-expressing tumors. [Mol Cancer Ther 2008;7(1):90 -100]

show abstract

Section: Discussionmentioning

confidence: 99%

Molecular mechanisms for the activity of PX-478, an antitumor inhibitor of the hypoxia-inducible factor-1α

Koh

Spivak-Kroizman

Venturini

et al. 2008

Molecular Cancer Therapeutics

266

188

View full text Add to dashboard Cite

show abstract

“…It is argued that NO blocks mitochondrial respiration, thereby leaving more oxygen available for PHDs to reactivate them. Alternatively, NO is supposed to directly activate PHD in vitro (Wang et al, 2002). Irrespective of underlying molecular mechanism the abovementioned suggestions center on reactivating PHD activity by NO under hypoxia to promote subsequent proteasomal degradation of HIF-1␣.…”

Section: Discussionmentioning

confidence: 99%

“…RNI attenuate hypoxia-induced HIF-1␣ accumulation, HIF-1 DNA-binding and HIF-1 transcriptional activation (Liu et al, 1998;Sogawa et al, 1998;Huang et al, 1999). Although mechanistic details remained unclear it has been proposed that NO, derived from sodium nitroprusside, enhanced the interaction between HIF-1␣ and pVHL in vitro through reactivation of PHD activity (Wang et al, 2002). More recently it was proposed that NO, by blocking cytochrome c oxidase, leaves more oxygen available for PHD to regain its activity under hypoxic conditions (Hagen et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Calpain Mediates a von Hippel-Lindau Protein–independent Destruction of Hypoxia-inducible Factor-1α

Zhou

Köhl

Herr

et al. 2006

MBoC

View full text Add to dashboard Cite

Hypoxia-inducible factor 1 (HIF-1) is controlled through stability regulation of its alpha subunit, which is expressed under hypoxia but degraded under normoxia. Degradation of HIF-1␣ requires association of the von Hippel Lindau protein (pVHL) to provoke ubiquitination followed by proteasomal digestion. Besides hypoxia, nitric oxide (NO) stabilizes HIF-1␣ under normoxia but destabilizes the protein under hypoxia. To understand the role of NO under hypoxia we made use of pVHL-deficient renal carcinoma cells (RCC4) that show a high steady state HIF-1␣ expression under normoxia. Exposing RCC4 cells to hypoxia in combination with the NO donor DETA-NO (2,2-(hydroxynitrosohydrazono) bis-ethanimine), but not hypoxia or DETA-NO alone, decreased HIF-1␣ protein and attenuated HIF-1 transactivation. Mechanistically, we noticed a role of calpain because calpain inhibitors reversed HIF-1␣ degradation. Furthermore, chelating intracellular calcium attenuated HIF-1␣ destruction by hypoxia/DETA-NO, whereas a calcium increase was sufficient to lower the amount of HIF-1␣ even under normoxia. An active role of calpain in lowering HIF-1␣ amount was also evident in pVHL-containing human embryonic kidney cells when the calcium pump inhibitor thapsigargin reduced HIF-1␣ that was stabilized by the prolyl hydroxylase inhibitor dimethyloxalylglycine (DMOG). We conclude that calcium contributes to HIF-1␣ destruction involving the calpain system. INTRODUCTIONThe transcription factor hypoxia-inducible factor-1 (HIF-1) constitutes a central component in coordinating adaptive responses toward low oxygen availability, i.e., hypoxia. HIF-1 is a heterodimer composed of the 120-kDa HIF-1␣ subunit and the 91-94-kDa HIF-1␤ subunit (Semenza and Wang, 1992;Wang and Semenza, 1995). Although HIF-1␤ is constitutively expressed, oxygen facilitates continues destruction of HIF-1␣ via the 26S proteasome. This requires polyubiquitination by an E3-ubiquitin ligase complex that contains the von Hippel Lindau protein (pVHL; Kaelin, 2002;Bruick, 2003;Huang and Bunn, 2003;Pugh and Ratcliffe, 2003;Semenza, 2003). Ubiquitination and binding of pVHL to HIF-1␣ demands hydroxylation of Pro564 and/or Pro402 within the oxygen-dependent degradation domain (ODD) of HIF-1␣ (Ivan et al., 2001;Jaakkola et al., 2001;Masson et al., 2001). Hydroxylation is mediated by prolyl hydroxylases, also known as PH domain-containing enzymes (PHD, i.e., PHD1 to PHD4; Bruick and McKnight, 2001;Epstein et al., 2001;Oehme et al., 2002). In addition to regulating HIF-1␣ protein stability, oxygen affects the transcriptional activity of HIF-1, by regulating hydroxylation of a critical Asn803 residue within the C-terminal transactivation domain (CTAD) of HIF-1␣ (Lando et al., 2002). The asparagine hydroxylase known as FIH (factor inhibiting HIF; Mahon et al., 2001;Hewitson et al., 2002) renders CTAD unable to bind the coactivator p300/CBP. Thus, hypoxia attenuates Pro564/402 as well as Asn803 modifications, provoking protein stabilization and coactivator recruitment, which concomitantly results...

show abstract

“…Our interest in these experiments was sparked by a report (Wang et al, 2002) postulating activation of HIF-1␣ prolyl hydroxylase activity in cytoplasmic extracts prepared from hypoxic cells.…”

Section: No Inhibits Hif-1␣ Prolyl Hydroxylasesmentioning

confidence: 99%

“…At present we do not have an explanation for the difference between normoxic and hypoxic NO effects. Although our study was in progress, it was reported that NO-evoked downregulation of HIF-1␣ in hypoxia is mediated by activation of PHDs (Wang et al, 2002) in cytoplasmic extracts containing partially purified PHDs. In our in vitro assay we failed to detect an activating impact of GSNO on recombinant PHDs in hypoxia.…”

mentioning

confidence: 93%

Nitric Oxide Impairs Normoxic Degradation of HIF-1α by Inhibition of Prolyl Hydroxylases

Metzen

Zhou

Jelkmann

et al. 2003

MBoC

379

261

View full text Add to dashboard Cite

Hypoxia inducible factor-1 (HIF-1) is the master regulator of metabolic adaptation to hypoxia. It is appreciated that HIF-1␣ accumulation is achieved under normoxic conditions by e.g., nitric oxide. We determined molecular mechanisms of HIF-1␣ accumulation under the impact of S-nitrosoglutathione (GSNO). In human embryonic kidney cells GSNO provoked nuclear accumulation of HIF-1␣. This appeared unrelated to gene transcription and protein translation, thus pointing to inhibition of HIF-1␣ degradation. Indeed, GSNO as well as the hypoxia mimic CoCl 2 decreased ubiquitination of HIF-1␣ and GSNO-induced HIF-1␣ failed to coimmunoprecipitate with pVHL (von Hippel Lindau protein). Considering that HIF-1␣-pVHL interactions require prolyl hydroxylation of HIF-1␣, we went on to demonstrate inhibition of HIF-1␣ prolyl hydroxylases (PHDs) by GSNO. In vitro HIF-1␣-pVHL interactions revealed that GSNO dose-dependently inhibits PHD activity but not the interaction of a synthetic peptide resembling the hydroxylated oxygen-dependent degradation domain of HIF-1␣ with pVHL. We conclude that GSNO-attenuated prolyl hydroxylase activity accounts for HIF-1␣ accumulation under conditions of NO formation during normoxia and that PHD activity is subject to regulation by NO. INTRODUCTIONThe heterodimeric transcription factor hypoxia inducible factor-1 (HIF-1) plays a central role in adaptation to decreased oxygen availability (Semenza, 2002;Wenger, 2002;Brü ne and Zhou, 2003). HIF-1 is composed of the two basic helix-loop-helix-Per-Arnt-Sim (bHLH-PAS) proteins HIF-1␣ and the aryl hydrocarbon receptor nuclear translocator (ARNT), also known as HIF-1␤ (Wang and Semenza, 1995). In many cell types, the mRNA of both HIF-1␣ and HIF-1␤ appear permanently expressed and HIF-1␤ protein is constitutively present. However, HIF-1␣ protein is kept at a low or undetectable level under normoxia. In hypoxia, HIF-1␣ is strikingly induced, translocates to the nucleus, and dimerizes with HIF-1␤ to form HIF-1, which binds to hypoxiaresponsive elements (HRE) in regulatory regions of an impressive array of target genes involved in angiogenesis, erythropoiesis, vasomotor control, and energy metabolism as well as in cell survival decisions (for references, see Maxwell et al., 2001;Wenger, 2002; Zhu et al., 2002). This makes HIF-1 the master regulator of oxygen homeostasis to meet cell and tissue requirements in a situation of oxygen deficiency.In normoxia HIF-1␣ is bound to the von Hippel Lindau protein (pVHL) (Maxwell et al., 1999;Hon et al., 2002;Min et al., 2002), which is the substrate recognizing component of an E3 ubiquitin ligase complex (Cockman et al., 2000;Ohh et al., 2000;Tanimoto et al., 2000). Consequently, HIF-1␣ is polyubiquitinated and degraded by the 26S proteasome system, thus accounting for its low normoxic level of expression (Salceda and Caro, 1997;Huang et al., 1998;Kallio et al., 1999). The requirement of pVHL for HIF-1␣ degradation is underscored in cells that do not contain a functional pVHL, which leads to high levels of HIF-1␣ in normox...

show abstract

HIF-1α-prolyl hydroxylase: molecular target of nitric oxide in the hypoxic signal transduction pathway

Cited by 61 publications

References 18 publications

Molecular mechanisms for the activity of PX-478, an antitumor inhibitor of the hypoxia-inducible factor-1α

Molecular mechanisms for the activity of PX-478, an antitumor inhibitor of the hypoxia-inducible factor-1α

Calpain Mediates a von Hippel-Lindau Protein–independent Destruction of Hypoxia-inducible Factor-1α

Nitric Oxide Impairs Normoxic Degradation of HIF-1α by Inhibition of Prolyl Hydroxylases

Contact Info

Product

Resources

About