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https://www.arca.fiocruz.br/handle/icict/27201
DNA POLYMERASE BETA FROM TRYPANOSOMA CRUZI IS INVOLVED IN KINETOPLAST DNA REPLICATION AND REPAIR OF OXIDATIVE LESIONS
Author
Reis, Bruno Luiz Fonseca Schamber
Nardelli, Sheila
Silva, Carlos Gustavo Régis
Campos, Priscila Carneiro
Cerqueira, Paula Gonçalves
Lima, Sabrina Almeida
Franco, Glória Regina
Macedo, Andrea Mara
Pena, Sergio Danilo Junho
Cazaux, Christophe
Hoffmann, Jean-Sébastien
Motta, Maria Cristina Machado
Schenkman, Sergio
Teixeira, Santuza Maria Ribeiro
Machado, Carlos Renato
Nardelli, Sheila
Silva, Carlos Gustavo Régis
Campos, Priscila Carneiro
Cerqueira, Paula Gonçalves
Lima, Sabrina Almeida
Franco, Glória Regina
Macedo, Andrea Mara
Pena, Sergio Danilo Junho
Cazaux, Christophe
Hoffmann, Jean-Sébastien
Motta, Maria Cristina Machado
Schenkman, Sergio
Teixeira, Santuza Maria Ribeiro
Machado, Carlos Renato
Affilliation
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil / School of Medical Sciences. Superior Learning and Development Center. Campina Grande, PB, Brazil
Federal University of São Paulo. Microbiology, Immunology and Parasitology Department. São Paulo, SP, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
University of Toulouse. Institute of Pharmacology and Structural Biology. Toulouse, France
University of Toulouse. Institute of Pharmacology and Structural Biology. Toulouse, France
Federal University of Rio de Janeiro. Institute of Biophysics Carlos Chagas Filho. Hertha Meyer Cellular Ultra structure Laboratory. Rio de Janeiro, RJ, Brazil
Federal University of São Paulo. Microbiology, Immunology and Parasitology Department. São Paulo, SP, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of São Paulo. Microbiology, Immunology and Parasitology Department. São Paulo, SP, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
University of Toulouse. Institute of Pharmacology and Structural Biology. Toulouse, France
University of Toulouse. Institute of Pharmacology and Structural Biology. Toulouse, France
Federal University of Rio de Janeiro. Institute of Biophysics Carlos Chagas Filho. Hertha Meyer Cellular Ultra structure Laboratory. Rio de Janeiro, RJ, Brazil
Federal University of São Paulo. Microbiology, Immunology and Parasitology Department. São Paulo, SP, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Federal University of Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil
Abstract
Specific DNA repair pathways from Trypanosoma cruzi are believed to protect genomic DNA and kinetoplast DNA (kDNA) from mutations. Particular pathways are supposed to operate in order to repair nucleotides oxidized by reactive oxygen species (ROS) during parasite infection, being 7,8-dihydro-8-oxoguanine (8oxoG) a frequent and highly mutagenic base alteration. If unrepaired, 8oxoG can lead to cytotoxic base transversions during DNA replication. In mammals, DNA polymerase beta (Polβ) is mainly involved in base excision repair (BER) of oxidative damage. However its biological role in T. cruzi is still unknown. We show, by immunofluorescence localization, that T. cruzi DNA polymerase beta (Tcpolβ) is restricted to the antipodal sites of kDNA in replicative epimastigote and amastigote developmental stages, being strictly localized to kDNA antipodal sites between G1/S and early G2 phase in replicative epimastigotes. Nevertheless, this polymerase was detected inside the mitochondrial matrix of trypomastigote forms, which are not able to replicate in culture. Parasites over expressing Tcpolβ showed reduced levels of 8oxoG in kDNA and an increased survival after treatment with hydrogen peroxide when compared to control cells. However, this resistance was lost after treating Tcpolβ overexpressors with methoxiamine, a potent BER inhibitor. Curiously, a presumed DNA repair focus containing Tcpolβ was identified in the vicinity of kDNA of cultured wild type epimastigotes after treatment with hydrogen peroxide. Taken together our data suggest participation of Tcpolβ during kDNA replication and repair of oxidative DNA damage induced by genotoxic stress in this organelle.
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