Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in Mice
Tumor hypoxia plays a crucial role in tumorigenesis. Under hypoxia, hypoxia-inducible factor 1α (HIF-1α) regulates activation of genes promoting malignant progression. Under normoxia, HIF-1α is hydroxylated on prolines 402 and 564 and is targeted for ubiquitin-mediated degradation by interacting wit...
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2008-05-01
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Series: | Molecular Imaging |
Online Access: | https://doi.org/10.2310/7290.2008.00017 |
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doaj-52906f6483cb4869a528cc85b47243232021-04-02T11:23:30ZengHindawi - SAGE PublishingMolecular Imaging1536-01212008-05-01710.2310/7290.2008.0001710.2310_7290.2008.00017Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in MiceClara Y.H. ChoiDenise A. ChanRamasamy PaulmuruganPatrick D. SutphinQuynh-Thu LeAlbert C. KoongWayne ZundelSanjiv S. GambhirAmato J. GiacciaTumor hypoxia plays a crucial role in tumorigenesis. Under hypoxia, hypoxia-inducible factor 1α (HIF-1α) regulates activation of genes promoting malignant progression. Under normoxia, HIF-1α is hydroxylated on prolines 402 and 564 and is targeted for ubiquitin-mediated degradation by interacting with the von Hippel-Lindau protein complex (pVHL). We have developed a novel method of studying the interaction between HIF-1α and pVHL using the split firefly luciferase complementation-based bioluminescence system in which HIF-1α and pVHL are fused to amino-terminal and carboxy-terminal fragments of the luciferase, respectively. We demonstrate that hydroxylation-dependent interaction between the HIF-1α and pVHL leads to complementation of the two luciferase fragments, resulting in bioluminescence in vitro and in vivo. Complementation-based bioluminescence is diminished when mutant pVHLs with decreased affinity for binding HIF-1α are used. This method represents a new approach for studying interaction of proteins involved in the regulation of protein degradation.https://doi.org/10.2310/7290.2008.00017 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Clara Y.H. Choi Denise A. Chan Ramasamy Paulmurugan Patrick D. Sutphin Quynh-Thu Le Albert C. Koong Wayne Zundel Sanjiv S. Gambhir Amato J. Giaccia |
spellingShingle |
Clara Y.H. Choi Denise A. Chan Ramasamy Paulmurugan Patrick D. Sutphin Quynh-Thu Le Albert C. Koong Wayne Zundel Sanjiv S. Gambhir Amato J. Giaccia Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in Mice Molecular Imaging |
author_facet |
Clara Y.H. Choi Denise A. Chan Ramasamy Paulmurugan Patrick D. Sutphin Quynh-Thu Le Albert C. Koong Wayne Zundel Sanjiv S. Gambhir Amato J. Giaccia |
author_sort |
Clara Y.H. Choi |
title |
Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in Mice |
title_short |
Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in Mice |
title_full |
Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in Mice |
title_fullStr |
Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in Mice |
title_full_unstemmed |
Molecular Imaging of Hypoxia-Inducible Factor 1α and von Hippel-Lindau Interaction in Mice |
title_sort |
molecular imaging of hypoxia-inducible factor 1α and von hippel-lindau interaction in mice |
publisher |
Hindawi - SAGE Publishing |
series |
Molecular Imaging |
issn |
1536-0121 |
publishDate |
2008-05-01 |
description |
Tumor hypoxia plays a crucial role in tumorigenesis. Under hypoxia, hypoxia-inducible factor 1α (HIF-1α) regulates activation of genes promoting malignant progression. Under normoxia, HIF-1α is hydroxylated on prolines 402 and 564 and is targeted for ubiquitin-mediated degradation by interacting with the von Hippel-Lindau protein complex (pVHL). We have developed a novel method of studying the interaction between HIF-1α and pVHL using the split firefly luciferase complementation-based bioluminescence system in which HIF-1α and pVHL are fused to amino-terminal and carboxy-terminal fragments of the luciferase, respectively. We demonstrate that hydroxylation-dependent interaction between the HIF-1α and pVHL leads to complementation of the two luciferase fragments, resulting in bioluminescence in vitro and in vivo. Complementation-based bioluminescence is diminished when mutant pVHLs with decreased affinity for binding HIF-1α are used. This method represents a new approach for studying interaction of proteins involved in the regulation of protein degradation. |
url |
https://doi.org/10.2310/7290.2008.00017 |
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