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

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 s...

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Bibliographic Details
Published in:PLoS neglected tropical diseases 2016-12, Vol.10 (12), p.e0005181-e0005181
Main Authors: Pavani, Raphael Souza, da Silva, Marcelo Santos, Fernandes, Carlos Alexandre Henrique, Morini, Flavia Souza, Araujo, Christiane Bezerra, Fontes, Marcos Roberto de Mattos, Sant'Anna, Osvaldo Augusto, Machado, Carlos Renato, Cano, Maria Isabel, Fragoso, Stenio Perdigão, Elias, Maria Carolina
Format: Article
Language:English
Subjects:
Animals
Biology and Life Sciences
Cell cycle
Cell Differentiation
Cell growth
Chagas Disease
Computer Simulation
Deoxyribonucleic acid
DNA
DNA binding proteins
DNA, Protozoan - metabolism
DNA, Single-Stranded - metabolism
Funding
Humans
Metabolism
Molecular Dynamics Simulation
Physical Sciences
Physiological aspects
Protein Binding
Proteins
Protozoa
Protozoan Proteins - chemistry
Protozoan Proteins - genetics
Protozoan Proteins - isolation & purification
Protozoan Proteins - metabolism
Replication Protein A - chemistry
Replication Protein A - genetics
Replication Protein A - isolation & purification
Replication Protein A - metabolism
Research and Analysis Methods
Simulation
Tropical diseases
Trypanosoma cruzi
Trypanosoma cruzi - genetics
Trypanosoma cruzi - physiology
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Summary: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.
ISSN:1935-2735
1935-2727
1935-2735
DOI:10.1371/journal.pntd.0005181