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Functional analysis of Ataxia-telangiectasia mutated (ATM) and Ataxia-telangiectasia and RAD3-related (ATR) in DNA damage response in Trypanosoma cruzi

Grant number: 14/21557-6
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): December 01, 2014
Effective date (End): June 30, 2016
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal Investigator:Maria Carolina Quartim Barbosa Elias Sabbaga
Grantee:Luis Eduardo Lazarim
Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
Associated research grant:13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling, AP.CEPID


The Trypanosoma cruzi cell cycle includes proliferative form and forms that are not able to proliferate. The signaling pathway that leads disruption of DNA replication in non-proliferating forms has not been elucidated. As other metabolic processes of DNA, replication and transcription are inactivated in non-proliferating form and as DNA damage can stop the cell cycle, we are interested in evaluating whether the signaling pathways of DNA damage are present and may be activated in non-proliferative forms of T. cruzi. Our group identified in T. cruzi genome (, ATM and ATR homologous to yeast genes that encode proteins known to be involved in the DNA damage signaling pathway. These proteins, once activated, can cause cell cycle arrest. In these genes, we identified PI3Kc_like super family domain in ATM homologue and PIKKc_ATR domain in ATR homologue. In this project, we intend to express part of the proteins encoded by these genes to obtain antibody against these proteins. Then, we intend to induce double strand breaks and proliferative stress in proliferative forms in order to check the role of these proteins in the DNA damage signaling. To do so, we will hold immunofluorescence assay to evaluate nuclear foci formation that correspond to activation of ATM and ATR. Once proven the role of these proteins in DNA damage signaling, we will evaluate the possible activation of these proteins in non-proliferative T. cruzi lifeforms. Functional characterization of ATM and ATR and the availability of these antibodies provide important tools for studying replication, repair and DNA damage response in Trypanosoma.

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