Replication Protein A Presents Canonical Functions and Is Also Involved in the Differentiation Capacity of Trypanosoma cruzi

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Pavani, Raphael Souza ; da Silva, Marcelo Santos ; Henrique Fernandes, Carlos Alexandre ; Morini, Flavia Souza ; Araujo, Christiane Bezerra ; de Mattos Fontes, Marcos Roberto ; Sant'Anna, Osvaldo Augusto ; Machado, Carlos Renato ; Cano, Maria Isabel ; Fragoso, Stenio Perdigao ; Elias, Maria Carolina

• Tipo de documento: Artigo Científico
• Fonte: PLoS Neglected Tropical Diseases; v. 10, n. 12 DEC 2016.
• Citações Web of Science: 5

Resumo

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. (AU)

• Processo FAPESP: 14/13375-5 - Origens da replicação em tripanossomas
• Beneficiário: Christiane Bezerra de Araujo
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/24170-5 - Dinâmica da replicação do DNA em Trypanosoma cruzi: caracterização do licenciamento e da taxa de replicação
• Beneficiário: Marcelo Santos da Silva
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/17864-8 - Estudos Estruturais de Fosfolipases A2 neurotoxicas
• Beneficiário: Carlos Alexandre Henrique Fernandes
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/02978-0 - Análise funcional do complexo RPA em Trypanosoma cruzi e seu envolvimento com DNA telomérico
• Beneficiário: Raphael Souza Pavani
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 13/07467-1 - CeTICS - Centro de Toxinas, Imuno-Resposta e Sinalização Celular
• Beneficiário: Hugo Aguirre Armelin
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 15/10580-0 - Caracterização de checkpoint intra-S em células de Trypanosoma
• Beneficiário: Maria Carolina Quartim Barbosa Elias Sabbaga
• Modalidade de apoio: Auxílio à Pesquisa - Regular