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Transcriptional Regulation of Genes via Hypoxia-Inducible Factor

  • Olga Roche
  • Michael OhhEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 809)

Abstract

Hypoxia-inducible factor (HIF) is the principal transcription factor that regulates adaptive physiologic responses to compromised oxygen tension. von Hippel–Lindau (VHL) tumor-suppressor protein binds and ubiquitylates the catalytic α subunit of HIF in an oxygen-dependent manner for rapid destruction via the 26S proteasome, thereby establishing VHL as a critical negative regulator of HIF. Mutations in VHL cause VHL disease, which is frequently characterized by the overexpression of HIFα and the development of tumors in multiple organ systems, including the central nervous system and the kidney. Here, we describe classical experimental approaches to demonstrate and validate HIF-responsive transcriptional regulation of genes.

Key words

Hypoxia HIF HRE EMSA Real-time PCR 

Notes

Acknowledgments

This work was supported by grants from the Canadian Institutes of Health Research (CIHR; MOP77718) and the Canadian Cancer Society (16056 and 18460). O. Roche is a recipient of a CIHR postdoctoral fellowship. M. Ohh is a Canada Research Chair.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Laboratory Medicine and PathobiologyUniversity of TorontoTorontoCanada

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