Bacterially produced human HIF-1alpha is competent for heterodimerization and specific DNA-binding.
Hypoxia-inducible factor 1alpha (HIF-1alpha) is the regulatory subunit of HIF-1, the transcriptional activator and key mediator of the cellular response to hypoxia. Regulation of HIF-1alpha occurs at multiple levels and involves several different post-translational modifications. In order to examine the importance of these modifications for the basic function of HIF-1alpha, ... we have produced in bacteria recombinant full-length human HIF-1alpha using different expression systems. We show that this unmodified form of HIF-1alpha is able to form a stable heterodimer with the second subunit of HIF-1 (HIF-1beta or ARNT) when both proteins are co-expressed in Escherichia coli. Furthermore, this bacterially reconstituted heterodimer exhibits specific DNA-binding activity. These data indicate that post-translational modification of HIF-1alpha is not essential for its interaction with ARNT and DNA, and provide an in vitro system for the characterization of the molecular properties of HIF-1alpha.
Mesh Terms:
Aryl Hydrocarbon Receptor Nuclear Translocator, DNA, DNA-Binding Proteins, Dimerization, Escherichia coli, Gene Expression, Humans, Hypoxia-Inducible Factor 1, alpha Subunit, Protein Binding, Protein Structure, Quaternary, Receptors, Aryl Hydrocarbon, Recombinant Proteins, Solubility, Substrate Specificity, Transcription Factors
Aryl Hydrocarbon Receptor Nuclear Translocator, DNA, DNA-Binding Proteins, Dimerization, Escherichia coli, Gene Expression, Humans, Hypoxia-Inducible Factor 1, alpha Subunit, Protein Binding, Protein Structure, Quaternary, Receptors, Aryl Hydrocarbon, Recombinant Proteins, Solubility, Substrate Specificity, Transcription Factors
Biochem. Biophys. Res. Commun.
Date: Jun. 03, 2005
PubMed ID: 15850782
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