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Characterization of protein quinases involved in the translation control, metabolism and extracellular vesicles secretion in trypanosomatides

Grant number: 21/12883-0
Support Opportunities:Scholarships in Brazil - Post-Doctoral
Effective date (Start): December 01, 2021
Effective date (End): November 30, 2024
Field of knowledge:Biological Sciences - Biochemistry - Biochemistry of Microorganisms
Principal Investigator:Sergio Schenkman
Grantee:Nadjania Saraiva de Lira Silva
Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
Associated research grant:20/07870-4 - Mechanisms of trypanosomatid adaptation to hosts through the control of transcription, protein synthesis and secretion of extracellular vesicles, AP.TEM

Abstract

The main objective of this project and which will be developed by the postdoctoral student in this period will be related to the control of translation and formation of RNA granules and the relationship of these processes with the secretion of extracellular vesicles by Trypanosomes. For this, we intend to: 1) generate parasites containing TcK1, K2 and K3 Tags generated by CRISPR/Cas9. The parasites will be used for localization of these proteins in parasites subjected to different stresses and for experiments if co-immunoprecipitation to identify their substrates and regulators; 2) detect the formation and composition of RNA granules (proteins and RNAs) from parasites labeled in described proteins and in the knockouts of TcK1, TcK2 and TcK3 superexpressors; 3) check how TcK1, TcK2 knockouts affect the secretion of extracellular vesicles; 4) generate protein mutants involved in the secretion pathways of T. cruzi systematically and analyze the effects on the processes of endocytosis and secretion of vesicles in the different stages of T. Cruzi; 5) detect changes in the total proteome and surface of the parasites with the different mutations; 6) characterize as extracellular vesicles released in all cases. (AU)

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