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Repertoire, genealogy and genomic organization of cruzipain and homologous genes in Trypanosoma cruzi, T. cruzi-like and other trypanosome species.

Lima L, Ortiz PA, da Silva FM, Alves JM, Serrano MG, Cortez AP, Alfieri SC, Buck GA, Teixeira MM - PLoS ONE (2012)

Bottom Line: In this study, we compared 80 sequences of genes encoding cruzipain from 25 T. cruzi isolates representative of all discrete typing units (DTUs TcI-TcVI) and the new genotype Tcbat and 10 sequences of homologous genes from other species.Relatively homogeneous sequences are found within and among isolates of the same DTU except TcV and TcVI, which displayed sequences unique or identical to those of TcII and TcIII, supporting their origin from the hybridization between these two DTUs.Our findings corroborate cruzipain as valuable target for drugs, vaccine, diagnostic and genotyping approaches.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Parasitologia, ICB, Universidade de São Paulo, São Paulo, São Paulo, Brasil.

ABSTRACT
Trypanosoma cruzi, the agent of Chagas disease, is a complex of genetically diverse isolates highly phylogenetically related to T. cruzi-like species, Trypanosoma cruzi marinkellei and Trypanosoma dionisii, all sharing morphology of blood and culture forms and development within cells. However, they differ in hosts, vectors and pathogenicity: T. cruzi is a human pathogen infective to virtually all mammals whilst the other two species are non-pathogenic and bat restricted. Previous studies suggest that variations in expression levels and genetic diversity of cruzipain, the major isoform of cathepsin L-like (CATL) enzymes of T. cruzi, correlate with levels of cellular invasion, differentiation, virulence and pathogenicity of distinct strains. In this study, we compared 80 sequences of genes encoding cruzipain from 25 T. cruzi isolates representative of all discrete typing units (DTUs TcI-TcVI) and the new genotype Tcbat and 10 sequences of homologous genes from other species. The catalytic domain repertoires diverged according to DTUs and trypanosome species. Relatively homogeneous sequences are found within and among isolates of the same DTU except TcV and TcVI, which displayed sequences unique or identical to those of TcII and TcIII, supporting their origin from the hybridization between these two DTUs. In network genealogies, sequences from T. cruzi clustered tightly together and closer to T. c. marinkellei than to T. dionisii and largely differed from homologues of T. rangeli and T. b. brucei. Here, analysis of isolates representative of the overall biological and genetic diversity of T. cruzi and closest T. cruzi-like species evidenced DTU- and species-specific polymorphisms corroborating phylogenetic relationships inferred with other genes. Comparison of both phylogenetically close and distant trypanosomes is valuable to understand host-parasite interactions, virulence and pathogenicity. Our findings corroborate cruzipain as valuable target for drugs, vaccine, diagnostic and genotyping approaches.

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Network genealogies of predicted amino acid sequences from all domains of genes encoding cruzipain in T. cruzi and homologues in other trypanosome species.Networks produced using the Neighbour-Net algorithm in SplitsTree v4.11.3, excluding all conserved sites and with Uncorrected p-distance. Networks were produced using entire sequences (A), pre- and pro-domains (B) or restricted to catalytic domains (C) of cruzipain encoding genes from the different trypanosomes are indicated by different symbols and colors according to the legend. Numbers in nodes correspond to support values estimated by performing 100 bootstrap replicates using the same parameter optimized for network inferences.
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pone-0038385-g002: Network genealogies of predicted amino acid sequences from all domains of genes encoding cruzipain in T. cruzi and homologues in other trypanosome species.Networks produced using the Neighbour-Net algorithm in SplitsTree v4.11.3, excluding all conserved sites and with Uncorrected p-distance. Networks were produced using entire sequences (A), pre- and pro-domains (B) or restricted to catalytic domains (C) of cruzipain encoding genes from the different trypanosomes are indicated by different symbols and colors according to the legend. Numbers in nodes correspond to support values estimated by performing 100 bootstrap replicates using the same parameter optimized for network inferences.

Mentions: Cruzipain amino acid sequences from T. cruzi isolates were relatively conserved in all domains. The ratio of non-synonymous (dN) to synonymous (dS) substitutions in the catalytic domain was dN/dS <1 by comparing the distinct DTUs of T. cruzi and T. cruzi-like species, suggesting that the enzymatic domain of cruzipain genes has been subjected to stabilizing selection for the conservation of metabolic function within the subgenus Schizotrypanum. Sequences encoding homologous cruzipain genes of T. c. marinkellei were closely related to those of T. cruzi (∼6.5% divergence) but the divergences were larger than those separating the T. cruzi DTUs (maximum of ∼2.5%). Sequences from T. cruzi largely diverged from homologues of T. dionisii (∼20%), T. rangeli (∼33%) and T. b. brucei (∼43%) in all domains (Fig.1). Genealogies based on whole cruzipain genes, or restricted to pre-pro or to catalytic domains, resulted in identical topologies (Fig. 2).


Repertoire, genealogy and genomic organization of cruzipain and homologous genes in Trypanosoma cruzi, T. cruzi-like and other trypanosome species.

Lima L, Ortiz PA, da Silva FM, Alves JM, Serrano MG, Cortez AP, Alfieri SC, Buck GA, Teixeira MM - PLoS ONE (2012)

Network genealogies of predicted amino acid sequences from all domains of genes encoding cruzipain in T. cruzi and homologues in other trypanosome species.Networks produced using the Neighbour-Net algorithm in SplitsTree v4.11.3, excluding all conserved sites and with Uncorrected p-distance. Networks were produced using entire sequences (A), pre- and pro-domains (B) or restricted to catalytic domains (C) of cruzipain encoding genes from the different trypanosomes are indicated by different symbols and colors according to the legend. Numbers in nodes correspond to support values estimated by performing 100 bootstrap replicates using the same parameter optimized for network inferences.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3369871&req=5

pone-0038385-g002: Network genealogies of predicted amino acid sequences from all domains of genes encoding cruzipain in T. cruzi and homologues in other trypanosome species.Networks produced using the Neighbour-Net algorithm in SplitsTree v4.11.3, excluding all conserved sites and with Uncorrected p-distance. Networks were produced using entire sequences (A), pre- and pro-domains (B) or restricted to catalytic domains (C) of cruzipain encoding genes from the different trypanosomes are indicated by different symbols and colors according to the legend. Numbers in nodes correspond to support values estimated by performing 100 bootstrap replicates using the same parameter optimized for network inferences.
Mentions: Cruzipain amino acid sequences from T. cruzi isolates were relatively conserved in all domains. The ratio of non-synonymous (dN) to synonymous (dS) substitutions in the catalytic domain was dN/dS <1 by comparing the distinct DTUs of T. cruzi and T. cruzi-like species, suggesting that the enzymatic domain of cruzipain genes has been subjected to stabilizing selection for the conservation of metabolic function within the subgenus Schizotrypanum. Sequences encoding homologous cruzipain genes of T. c. marinkellei were closely related to those of T. cruzi (∼6.5% divergence) but the divergences were larger than those separating the T. cruzi DTUs (maximum of ∼2.5%). Sequences from T. cruzi largely diverged from homologues of T. dionisii (∼20%), T. rangeli (∼33%) and T. b. brucei (∼43%) in all domains (Fig.1). Genealogies based on whole cruzipain genes, or restricted to pre-pro or to catalytic domains, resulted in identical topologies (Fig. 2).

Bottom Line: In this study, we compared 80 sequences of genes encoding cruzipain from 25 T. cruzi isolates representative of all discrete typing units (DTUs TcI-TcVI) and the new genotype Tcbat and 10 sequences of homologous genes from other species.Relatively homogeneous sequences are found within and among isolates of the same DTU except TcV and TcVI, which displayed sequences unique or identical to those of TcII and TcIII, supporting their origin from the hybridization between these two DTUs.Our findings corroborate cruzipain as valuable target for drugs, vaccine, diagnostic and genotyping approaches.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Parasitologia, ICB, Universidade de São Paulo, São Paulo, São Paulo, Brasil.

ABSTRACT
Trypanosoma cruzi, the agent of Chagas disease, is a complex of genetically diverse isolates highly phylogenetically related to T. cruzi-like species, Trypanosoma cruzi marinkellei and Trypanosoma dionisii, all sharing morphology of blood and culture forms and development within cells. However, they differ in hosts, vectors and pathogenicity: T. cruzi is a human pathogen infective to virtually all mammals whilst the other two species are non-pathogenic and bat restricted. Previous studies suggest that variations in expression levels and genetic diversity of cruzipain, the major isoform of cathepsin L-like (CATL) enzymes of T. cruzi, correlate with levels of cellular invasion, differentiation, virulence and pathogenicity of distinct strains. In this study, we compared 80 sequences of genes encoding cruzipain from 25 T. cruzi isolates representative of all discrete typing units (DTUs TcI-TcVI) and the new genotype Tcbat and 10 sequences of homologous genes from other species. The catalytic domain repertoires diverged according to DTUs and trypanosome species. Relatively homogeneous sequences are found within and among isolates of the same DTU except TcV and TcVI, which displayed sequences unique or identical to those of TcII and TcIII, supporting their origin from the hybridization between these two DTUs. In network genealogies, sequences from T. cruzi clustered tightly together and closer to T. c. marinkellei than to T. dionisii and largely differed from homologues of T. rangeli and T. b. brucei. Here, analysis of isolates representative of the overall biological and genetic diversity of T. cruzi and closest T. cruzi-like species evidenced DTU- and species-specific polymorphisms corroborating phylogenetic relationships inferred with other genes. Comparison of both phylogenetically close and distant trypanosomes is valuable to understand host-parasite interactions, virulence and pathogenicity. Our findings corroborate cruzipain as valuable target for drugs, vaccine, diagnostic and genotyping approaches.

Show MeSH
Related in: MedlinePlus