Investigation of molecular chaperones cell expression and their function by complementation in knockout yeasts

• Grant number: 15/08909-3
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): September 01, 2015
• Effective date (End): April 30, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Carlos Henrique Inacio Ramos
• Grantee: Fernando Diez García
• Home Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 12/50161-8 - Study of the structure and function of the Hsp90 chaperone with emphasis on its role in cellular homeostasis, AP.TEM

Abstract

The main objective of the Tematico is to characterize the structure and function of the Hsp90/Hsp70 chaperone system (chaperones and its co-chaperones). For that we are going to pufirify heterologous proteins from various organisms, including human, sugarcane and Leishmania. A major effort to characterize the function of these proteins has been done through the implementation of cells in tests to validate and extend the knowledge gained through functional studies in vitro. In this sense we have successfully initiated the investigation of several chaperones in yeast knockout strains of certain genes. At the moment we are studying the complementation in yeast Hsp104 knockout of genes (gene complementation from human and sugar cane) and the co-chaperone p23 (gene complementation from human and Leishmania). The study goes through phenotype verification steps (mainly resistance to stresses of thermal or chemical type), expression of the yeast chaperone and eventually construction of mutants to further studies (already we have some human constructs to initiate studies). In the future, in collaboration with experts in mass spectrometry we will try to investigate the effects on the proteome. Ideally, it would be interesting that the candidate could also perform qRT-PCR experiments to optimize validation purposes. It is expected that the results obtained can not only provide information about the interactions between chaperones and between them and other proteins.