Molecular characterization of Cyclophilin (TcCyP19) in Trypanosoma cruzi populations susceptible and resistant to benznidazole

Abstract

Cyclophilin (CyP), a peptidyl-prolyl cis/trans isomerase, is a key molecule with diverse biological functions that include roles in molecular chaperoning, stress response, immune modulation, and signal transduction. In this respect, CyP could serve as a potential drug target in disease-causing parasites. Previous studies employing proteomics techniques have shown that the TcCyP19 isoform was more abundant in a benznidazole (BZ)-resistant Trypanosoma cruzi population than in its susceptible counterpart. In this study, TcCyP19 has been characterized in BZ-susceptible and BZ-resistant T. cruzi populations. Phylogenetic analysis revealed a clear dichotomy between Cyphophilin A (CyPA) sequences from trypanosomatids and mammals. Sequencing analysis revealed that the amino acid sequences of TcCyP19 were identical among the T. cruzi samples analyzed. Southern blot analysis showed that TcCyP19 is a single-copy gene, located in chromosomal bands varying in size from 0.68 to 2.2 Mb, depending on the strain of T. cruzi. Northern blot and qPCR indicated that the levels of TcCyP19 mRNA were twofold higher in drug-resistant T. cruzi populations than in their drug-susceptible counterparts. Similarly, as determined by two-dimensional gel electrophoresis immunoblot, the expression of TcCyP19 protein was increased to the same degree in BZ-resistant T. cruzi populations. No differences in TcCyP19 mRNA and protein expression levels were observed between the susceptible and the naturally resistant T. cruzi strains analyzed. Taken together, these data indicate that cyclophilin TcCyP19 expression is up-regulated at both transcriptional and translational levels in T. cruzi populations that were in vitro-induced and in vivo-selected for resistance to BZ.

Transcriptome and Functional Genomics Reveal the Participation of Adenine Phosphoribosyltransferase in Trypanosoma cruzi Resistance to Benznidazole

“…It has been proposed that this gene may contribute to counteracting the chemical stress stimulus through mechanisms such as increasing protein‐folding activity, which is necessary for the efficient expression of functional polypeptides. This suggests that cyclophilin plays an important biological role in resistance to Bz [Rego et al, ]. Other genes implicated in transport, such as hexose transporters [Murta et al, ] and ABC transporters [Zingales et al, ], have been linked to resistance in T. cruzi ; consistent with our results, these have also been found to be up‐regulated.…”

“…Our results coincide with the findings of previous studies, which have shown this gene to be up‐regulated at the transcriptional and translational levels in Bz‐resistant T . cruzi [Andrade et al, ; Rego et al, ]. It has been proposed that this gene may contribute to counteracting the chemical stress stimulus through mechanisms such as increasing protein‐folding activity, which is necessary for the efficient expression of functional polypeptides.…”

Cyclophilin 19 secreted in the host cell cytosol byTrypanosoma cruzipromotesROSproduction required for parasite growth

“…Tc Cyp19 is 71.9% identical to human CypA (Bua et al, 2001), and inhibitors of cyclophilins such as cyclosporin A and its analogs reduce epimastigote proliferation and cellular infection by trypomastigotes (Bua et al, 2008; Bua, Ruiz, Potenza, & Fichera, 2004; Carraro, Bua, Ruiz, & Paulino, 2007). Tc Cyp19 is involved in the oxidant/antioxidant responses of T. cruzi (Bustos, Perrone, Milduberger, Postan, & Bua, 2015) and parasites selected by resistance to benznidazole, which acts through oxidative damage, show increased Tc Cyp19 expression (Rego et al, 2015).…”

A Functional Analysis of the Cyclophilin Repertoire in the Protozoan Parasite Trypanosoma Cruzi

“…Besides, the protein expression and mRNA transcription of the T. cruzi cyclophilin of 19 kDa were twofold higher in those parasites populations who were in vitro induced to be resistant to the parasiticidal drug benznidazol. The chaperoning activity of the overexpressed Tc CyP19 could be associated with the activity of enzymes with antioxidant defense conferring the parasite resistance to benznidazol [50].…”