Autor(es): Pavani, Raphael Souza ; da Silva, Marcelo Santos ; Fernandes, Carlos Alexandre Henrique [UNESP] ; Morini, Flavia Souza ; Araujo, Christiane Bezerra ; Fontes, Marcos Roberto de Mattos [UNESP] ; Sant’Anna, Osvaldo Augusto ; Machado, Carlos Renato ; Cano, Maria Isabel [UNESP] ; Fragoso, Stenio Perdigão ; Elias, Maria Carolina
Data: 2018
Identificador Persistente: http://hdl.handle.net/11449/174059
Origem: Oasisbr
Made available in DSpace on 2018-12-11T17:08:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-12-16
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Replication Protein A (RPA), the major single stranded DNA binding protein in eukaryotes, is composed of three subunits and is a fundamental player in DNA metabolism, participating in replication, transcription, repair, and the DNA damage response. In human pathogenic trypanosomatids, only limited studies have been performed on RPA-1 from Leishmania. Here, we performed in silico, in vitro and in vivo analysis of Trypanosoma cruzi RPA-1 and RPA-2 subunits. Although computational analysis suggests similarities in DNA binding and Ob-fold structures of RPA from T. cruzi compared with mammalian and fungi RPA, the predicted tridimensional structures of T. cruzi RPA-1 and RPA-2 indicated that these molecules present a more flexible tertiary structure, suggesting that T. cruzi RPA could be involved in additional responses. Here, we demonstrate experimentally that the T. cruzi RPA complex interacts with DNA via RPA-1 and is directly related to canonical functions, such as DNA replication and DNA damage response. Accordingly, a reduction of TcRPA-2 expression by generating heterozygous knockout cells impaired cell growth, slowing down S-phase progression. Moreover, heterozygous knockout cells presented a better efficiency in differentiation from epimastigote to metacyclic trypomastigote forms and metacyclic trypomastigote infection. Taken together, these findings indicate the involvement of TcRPA in the metacyclogenesis process and suggest that a delay in cell cycle progression could be linked with differentiation in T. cruzi.
Laboratório Especial de Ciclo Celular Instituto Butantan São Paulo
Center of Toxins Immune Response and Cell Signaling—CeTICS Instituto Butantan São Paulo
Departamento de Física e Biofísica Instituto de Biociências Universidade Estadual Paulista Júlio de Mesquita Filho -UNESP Botucatu
Instituto Carlos Chagas Fiocruz-PR
Laboratório de Imunoquímica Instituto Butantan São Paulo
Departamento de Bioquímica e Imunologia ICB Universidade Federal de Minas Gerais
Departamento de Genética Instituto de Biociências Universidade Estadual Paulista Julio Mesquita Filho—UNESP Botucatu
Departamento de Física e Biofísica Instituto de Biociências Universidade Estadual Paulista Júlio de Mesquita Filho -UNESP Botucatu
Departamento de Genética Instituto de Biociências Universidade Estadual Paulista Julio Mesquita Filho—UNESP Botucatu
FAPESP: 2013/07467-1
FAPESP: 2013/17864-8
FAPESP: 2014/02978-0
FAPESP: 2014/13375-5
FAPESP: 2014/24170-5
FAPESP: 2015/10508-0
