Bacterially produced human HIF-1α is competent for heterodimerization and specific DNA-binding

Hypoxia-inducible factor 1α (HIF-1α) is the regulatory subunit of HIF-1, the transcriptional activator and key mediator of the cellular response to hypoxia. Regulation of HIF-1α occurs at multiple levels and involves several different post-translational modifications. In order to examine the importa...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2005-06, Vol.331 (2), p.464-470
Main Authors: Chachami, Georgia, Paraskeva, Efrosyni, Georgatsou, Eleni, Bonanou, Sofia, Simos, George
Format: Article
Language:English
Subjects:
ARNT
Bacterial expression
DNA-binding
Escherichia coli
Heterodimerization
HIF-1α
Hypoxia-inducible factor 1
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Summary:Hypoxia-inducible factor 1α (HIF-1α) is the regulatory subunit of HIF-1, the transcriptional activator and key mediator of the cellular response to hypoxia. Regulation of HIF-1α 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-1α, we have produced in bacteria recombinant full-length human HIF-1α using different expression systems. We show that this unmodified form of HIF-1α is able to form a stable heterodimer with the second subunit of HIF-1 (HIF-1β 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-1α 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-1α.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2005.03.193