title: 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.
authors.affiliation: Campus Amilcar Ferreira Sobral, Universidade Federal do Piauí - UFPI, Floriano, PI, Brazil; Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisa René Rachou - CPqRR/FIOCRUZ, Belo Horizonte, MG, Brazil; Instituto de Ciências do Mar, Universidade Federal do Ceará-Labomar/UFC, Fortaleza, CE, Brazil.
authors.forename: Juciane Vaz
authors.initials: JV
authors.lastname: Rêgo
authors.affiliation: Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisa René Rachou - CPqRR/FIOCRUZ, Belo Horizonte, MG, Brazil.
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Duarte
authors.affiliation: Campus Petrônio Portela, Universidade Federal do Piauí - UFPI,Teresina, PI, Brazil.
authors.forename: Daniel Barbosa
authors.initials: DB
authors.lastname: Liarte
authors.affiliation: Campus Amilcar Ferreira Sobral, Universidade Federal do Piauí - UFPI, Floriano, PI, Brazil.
authors.forename: Francirlene
authors.initials: F
authors.lastname: de Carvalho Sousa
authors.affiliation: Campus Petrônio Portela, Universidade Federal do Piauí - UFPI,Teresina, PI, Brazil.
authors.forename: Humberto Medeiros
authors.initials: HM
authors.lastname: Barreto
authors.affiliation: Instituto Nacional de Parasitología "Dr. M. Fatala Chabén", A.N.L.I.S., Buenos Aires, Argentina.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Bua
authors.affiliation: Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisa René Rachou - CPqRR/FIOCRUZ, Belo Horizonte, MG, Brazil.
authors.forename: Alvaro José
authors.initials: AJ
authors.lastname: Romanha
authors.affiliation: Instituto de Ciências do Mar, Universidade Federal do Ceará-Labomar/UFC, Fortaleza, CE, Brazil.
authors.forename: Gandhi
authors.initials: G
authors.lastname: Rádis-Baptista
authors.affiliation: Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisa René Rachou - CPqRR/FIOCRUZ, Belo Horizonte, MG, Brazil. Electronic address: silvane@cpqrr.fiocruz.br.
authors.forename: Silvane Maria Fonseca
authors.initials: SM
authors.lastname: Murta
mesh_terms: D000595:Amino Acid Sequence; D002874:Chromosome Mapping; D021983:Cyclophilins / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D016054:DNA, Protozoan / Q000032:analysis / Q000737:chemistry; D004351:Drug Resistance; D018628:Gene Dosage; D005786:Gene Expression Regulation; D018503:Genome, Protozoan; D009593:Nitroimidazoles / Q000494:pharmacology*; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D016053:RNA, Protozoan / Q000032:analysis / Q000737:chemistry; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000737:chemistry / Q000187:drug effects / Q000235:genetics*
chemical_list: D016054:DNA, Protozoan; D009593:Nitroimidazoles; D012333:RNA, Messenger; D016053:RNA, Protozoan; D014344:Trypanocidal Agents; C431607:cyclophilin 19; D021983:Cyclophilins; C009999:benzonidazole
keywords: Benznidazole; Cyclophilin; Drug resistance; Trypanosoma cruzi