Structural studies of the complexes formed between the oxygen-dependent degradation domain (ODD) of HIF-1alpha and prolil-hydroxilases

• Grant number: 12/05997-0
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: August 01, 2012
• End date: January 31, 2013
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Andre Luis Berteli Ambrosio
• Grantee: Angela Maria Fala
• Supervisor: Grazyna Kochan
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Institution abroad: University of Oxford, England
• Associated to the scholarship: 10/13739-6 - Establishing heterologous expression and purification conditions of the human hypoxia-inducible factor 1, HIF-1, for crystallographic studies., BP.MS

Abstract

Cancer cells employ complex strategies to deal with oxygen deprivation, for instance by inducing the formation of new blood vessels and alterations in energy metabolism. Extensive work performed during the past decade has demonstrated the role of a hypoxia-inducible transcription factor, termed HIF, as master regulator of cellular adaptation to low oxygen. HIF-1 is a heterodimer, composed by and alpha and a beta subunit, that mediates adaptive responses to changes in tissue oxygenation, regulating the transcription of more than 100 known target genes. It has been extensively shown that the key to oxygen-dependent gene regulation by HIF is the alpha subunit, which is rapidly degraded under normoxic conditions via the proteasomal pathway and becomes stable under hypoxic conditions. However, the direct oxygen sensor that controls the fate of the alpha subunit, and therefore the whole HIF pathway, were identified as being members of a class of oxygen binding known as prolyl-hydrixilases, and are known as PHD1, 2 and 3. PHDs covalently modify HIF-1aplha (by hydroxilating two proline residues) thus enabling recognition by a multiprotein complex that tags it for rapid proteasomal degradation. A variety of human disorders, including cancer, are characterized by, or associated with, tissue hypoxia and manipulation of the PHD-HIF pathway via small molecules has been predicted to significantly alter their clinical course. Within this context, and according to both the current line of research of our lab and the project of the candidate and current MSc student Ângela Maria Fala, the proposed project aims structural studies of the complexes formed between the Oxygen-dependent degradation domain (ODD) of HIF-1 alpha and prolil-hydroxilases, in collaboration with the Biotecnology group of the Structural Genomics Consortium, at the University of Oxford, UK. The SGC is a leading global consortium focused on structural biology of human proteins with an interest in health. The internship will be supervised by Dr. Grazyna Kochan, who is responsible for the group, due to its extensive experience with structural studies of HIF and PHDs. In case of success in obtaining crystals of the complexes, the student will have access to the beamlines dedicated to macromolecular crystallography at Diamond Light Source, located in the vicinity of Oxford. (AU)