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Determining physical interactions required for R2TP complex assembly using biophysical and biochemical approaches

Grant number: 16/01603-9
Support type:Scholarships abroad - Research Internship - Scientific Initiation
Effective date (Start): June 01, 2016
Effective date (End): September 30, 2016
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal researcher:Lisandra Marques Gava Borges
Grantee:Carolina Colleti
Supervisor abroad: Walid A. Houry
Home Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
Research place: University of Toronto (U of T), Canada  
Associated to the scholarship:14/15432-6 - Effect of catechin EGCG in assembly of Hsp90 complex: co-chaperones, BP.IC

Abstract

Hsp90 is a highly conserved, widely distributed and essential molecular chaperone involved in protein homeostasis. Due to the high number of interactors or client proteins (over 600, including oncoproteins), Hsp90 works as a cellular hub and is considered a valid target for caner therapeutics. In 2005, Dr Houry's group discovered an Hsp90-interacting complex, which they named the R2TP complex. This complex is central for cell physiology, modulating pathways involved in cell cycle and growth, ribosome biogenesis and genome maintenance. The human R2TP complex is formed by the proteins Rvb1, Rvb2, PIH1D1 and RPAP3, with RPAP3 mediating the interaction between the R2TP complex and Hsp90. Hsp90 is essential for the stability and function of the R2TP complex. However, the exact mechanism by which the human Hsp90-R2TP interact with each other is poorly understood. In this project we propose to address this question by studying the interaction between Hsp90 and RPAP3 using surface plasmon resonance and isothermal titration calorimetry techniques. Mapping of the interaction interfaces between RPAP3 and PIH1D1 will also be carried out using analytical size exclusion chromatography and pull-down assays. The study of these complexes will be of great importance for understanding Hsp90-R2TP complex assembly and will help the candidate develop her skills in the field of protein biophysics and biochemistry. The knowledge acquired during the research internship in Toronto will develop her skills and will greatly aid in her studies in Brazil aimed at determining the effects of the ligands (-)-Epigallocatechin-3-gallate (EGCG) and curcumin on Hsp90 protein complexes. (AU)