Document Detail


Monitoring of cellular responses to hypoxia.
MedLine Citation:
PMID:  19957135     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Oxygen is essential for survival of aerobic organisms. Sensing changes in the environmental oxygen concentration and appropriate adaptation to such changes are essential for organisms to survive. Hypoxia inducible factor 1 (HIF-1) is the key transcription factor in controlling the expression of oxygen-dependent genes required for this adaptation. HIF-1 is a heterodimer of an oxygen dependent alpha-subunit and constitutive beta-subunit. Abundance and activity of HIF-1 is controlled by post-translational hydroxylation. Microscopic analysis of the assembly and activation process of HIF-1 has become an important tool to better understand HIF-1 regulation. Confocal laser microscopy provides exact images of HIF-1alpha that is translocated into the nucleus under hypoxia and its disappearance upon reoxygenation. To exactly follow the protein-protein interaction during the assembly process of HIF-1, both subunits were labeled by fusing them to fluorescent proteins. Fluorescence resonance energy transfer (FRET) was used to determine the interaction of both subunits in living cells by confocal microscopy.
Authors:
Christoph Wotzlaw; Joachim Fandrey
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  591     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2010  
Date Detail:
Created Date:  2009-12-03     Completed Date:  2010-02-18     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  243-55     Citation Subset:  IM    
Affiliation:
Institut f?r Physiologie, Universit?t Duisburg-Essen, Essen, Germany.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
Cell Hypoxia*
Cell Line, Tumor
Cell Nucleus / metabolism
Cell Survival
Fluorescence Resonance Energy Transfer
Fluorescent Antibody Technique
Hypoxia-Inducible Factor 1, alpha Subunit / chemistry,  metabolism
Mice
Microscopy, Confocal
Protein Multimerization
Protein Structure, Quaternary
Chemical
Reg. No./Substance:
0/Hypoxia-Inducible Factor 1, alpha Subunit

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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