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Defining post-translational modifications and interactors for Replication Protein A (RPA) from trypanosomatids associated with DNA damage.

Grant number: 16/24255-6
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): March 15, 2017
Effective date (End): November 14, 2017
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal researcher:Maria Carolina Quartim Barbosa Elias Sabbaga
Grantee:Raphael Souza Pavani
Supervisor abroad: Mark C Field
Home Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
Research place: University of Dundee, Scotland  
Associated to the scholarship:14/02978-0 - Functional analysis of RPA complex in Trypanosoma cruzi and its involvement with telomeric DNA, BP.DR

Abstract

Replication protein A (RPA) is a heterotrimeric complex involved in many aspects of DNA metabolism, such as replication and DNA damage responses (DDR). Post-translational modifications (PTM) are important for RPA regulation and to support the multiple functions that RPA mediates. In mammalian cells following DNA damage RPA is phosphorylated at multiple sites, and which serve to initiate a repair and checkpoint response. In trypanosomatids, although the RPA-1 homologue lacks the major domain involved in interaction with repair and checkpoint proteins in mammals, we have evidence that RPA is involved in DDR. Our previous data suggest that RPA is modified after DNA damage, but how this modification affects RPA function and what additional protein partners are required for the response remains unknown. In this project, we will use cryomilling technology followed by immunoprecipitation and mass spectrometry analysis to identify PTM and interactors of the RPA complex following DNA damage in Trypanosoma brucei. These new findings will contribute to elucidating the mechanisms and pathways that regulate parasite surveillance following genotoxic stress.

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