Replication Protein A presents canonical functions and is also involved in the differentiation capacity of Trypanosoma cruzi

Submitted by Luciane Willcox ([email protected]) on 2017-04-19T13:31:24Z No. of bitstreams: 1 Replication Protein A Presents Canonical Functions and Is Also Involved in the Differentiation Capacity of Trypanosoma cruzi.pdf: 2837768 bytes, checksum: 4bd66cfcd5f9128f11a7dd42e4e9ed77 (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2017-07-26T11:56:07Z (GMT) No. of bitstreams: 1 Replication Protein A Presents Canonical Functions and Is Also Involved in the Differentiation Capacity of Trypanosoma cruzi.pdf: 2837768 bytes, checksum: 4bd66cfcd5f9128f11a7dd42e4e9ed77 (MD5)Made available in DSpace on 2017-07-26T11:56:07Z (GMT). No. of bitstreams: 1 Replication Protein A Presents Canonical Functions and Is Also Involved in the Differentiation Capacity of Trypanosoma cruzi.pdf: 2837768 bytes, checksum: 4bd66cfcd5f9128f11a7dd42e4e9ed77 (MD5) Previous issue date: 2016Instituto Butantan. Laboratório Especial de Ciclo Celular. São Paulo, SP, Brasil. / Instituto Butantan. Center of Toxins. Immune Response and Cell Signaling—CeTICS. São Paulo, SP, Brazil.Instituto Butantan. Laboratório Especial de Ciclo Celular. São Paulo, SP, Brasil. / Instituto Butantan. Center of Toxins. Immune Response and Cell Signaling—CeTICS. São Paulo, SP, Brazil.Universidade Estadual Paulista Júlio de Mesquita Filho. Instituto de Biociências. Departamento de Física e Biofísica. Botucatu, SP, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Instituto Butantan. Laboratório Especial de Ciclo Celular. São Paulo, SP, Brasil. / Instituto Butantan. Center of Toxins. Immune Response and Cell Signaling—CeTICS. São Paulo, SP, Brazil.Universidade Estadual Paulista Júlio de Mesquita Filho. Instituto de Biociências. Departamento de Física e Biofísica. Botucatu, SP, Brasil.Instituto Butantan. Center of Toxins. Immune Response and Cell Signaling—CeTICS. São Paulo, SP, Brazil / Instituto Butantan. Laboratório de Imunoquímica. São Paulo, SP, Brasil.Universidade Federal de Minas Gerais. Departamento de Bioquímica e Imunologia. Belo Horizonte, MG, Brasil.Universidade Estadual Paulista Julio Mesquita Filho. Instituto de Biociências. Departamento de Genética. Botucatu, SP, Brasil.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