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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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Polymorphism and network analyses of catalytic domain sequences of cruzipain genes from T. cruzi isolates of TcI-VI and Tcbat.(A) Polymorphic nucleotide sites on catalytic domains of cruzipain encoding genes; (B) Network based on polymorphic nucleotides constructed with the Neighbour-Net algorithm excluding all conserved sites and with Uncorrected p-distance. The numbers in nodes correspond to bootstrap values from 100 replicates. CLBrener1-5 are sequences from TriTrypDB: Tc00.1047053509429.320, Tc00.1047053507537.20, Tc00.1047053507603.270, Tc00.1047053507603.260 and Tc00.1047053507537.10; *GenBank accession numbers of all sequences included in these analyses are listed on Table1. Major types of sequences from T. cruzi isolates of different DTUs are indicated by different colors according to the legend.
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pone-0038385-g004: Polymorphism and network analyses of catalytic domain sequences of cruzipain genes from T. cruzi isolates of TcI-VI and Tcbat.(A) Polymorphic nucleotide sites on catalytic domains of cruzipain encoding genes; (B) Network based on polymorphic nucleotides constructed with the Neighbour-Net algorithm excluding all conserved sites and with Uncorrected p-distance. The numbers in nodes correspond to bootstrap values from 100 replicates. CLBrener1-5 are sequences from TriTrypDB: Tc00.1047053509429.320, Tc00.1047053507537.20, Tc00.1047053507603.270, Tc00.1047053507603.260 and Tc00.1047053507537.10; *GenBank accession numbers of all sequences included in these analyses are listed on Table1. Major types of sequences from T. cruzi isolates of different DTUs are indicated by different colors according to the legend.

Mentions: Any attempt to associate cruzipain polymorphisms with biological features of T. cruzi requires a good appraisal of the diversity of gene copies within both one strain/isolate and each DTU. We have assessed the polymorphism on cruzipain gene copies by comparing 3 to 8 sequences from each isolate. Larger number of sequences (7–8) was analyzed from the isolates of hybrid DTUs TcV and TcVI (Table 1). Cruzipain gene copies (paralogous) from isolates of TcI, TcIII, TcIV and Tcbat were identical or highly similar in their amino acid sequences (Fig. 3), whereas diverged in 2 to 6 polymorphic sites in their nucleotide sequences (Fig. 4). We identified a total of 23 variant sequences of cruzipain sequences (Fig. 4). Relatively homogeneous but not identical copies were found by comparing sequences from 6 isolates of TcI and two of TcII. No polymorphic sites were found among sequences from 3 isolates of each TcIII and TcIV. Sequences from Non-Esmeraldo-like haplotype of CL Brener were identical to those of TcIII (Fig. 4).


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)

Polymorphism and network analyses of catalytic domain sequences of cruzipain genes from T. cruzi isolates of TcI-VI and Tcbat.(A) Polymorphic nucleotide sites on catalytic domains of cruzipain encoding genes; (B) Network based on polymorphic nucleotides constructed with the Neighbour-Net algorithm excluding all conserved sites and with Uncorrected p-distance. The numbers in nodes correspond to bootstrap values from 100 replicates. CLBrener1-5 are sequences from TriTrypDB: Tc00.1047053509429.320, Tc00.1047053507537.20, Tc00.1047053507603.270, Tc00.1047053507603.260 and Tc00.1047053507537.10; *GenBank accession numbers of all sequences included in these analyses are listed on Table1. Major types of sequences from T. cruzi isolates of different DTUs are indicated by different colors according to the legend.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038385-g004: Polymorphism and network analyses of catalytic domain sequences of cruzipain genes from T. cruzi isolates of TcI-VI and Tcbat.(A) Polymorphic nucleotide sites on catalytic domains of cruzipain encoding genes; (B) Network based on polymorphic nucleotides constructed with the Neighbour-Net algorithm excluding all conserved sites and with Uncorrected p-distance. The numbers in nodes correspond to bootstrap values from 100 replicates. CLBrener1-5 are sequences from TriTrypDB: Tc00.1047053509429.320, Tc00.1047053507537.20, Tc00.1047053507603.270, Tc00.1047053507603.260 and Tc00.1047053507537.10; *GenBank accession numbers of all sequences included in these analyses are listed on Table1. Major types of sequences from T. cruzi isolates of different DTUs are indicated by different colors according to the legend.
Mentions: Any attempt to associate cruzipain polymorphisms with biological features of T. cruzi requires a good appraisal of the diversity of gene copies within both one strain/isolate and each DTU. We have assessed the polymorphism on cruzipain gene copies by comparing 3 to 8 sequences from each isolate. Larger number of sequences (7–8) was analyzed from the isolates of hybrid DTUs TcV and TcVI (Table 1). Cruzipain gene copies (paralogous) from isolates of TcI, TcIII, TcIV and Tcbat were identical or highly similar in their amino acid sequences (Fig. 3), whereas diverged in 2 to 6 polymorphic sites in their nucleotide sequences (Fig. 4). We identified a total of 23 variant sequences of cruzipain sequences (Fig. 4). Relatively homogeneous but not identical copies were found by comparing sequences from 6 isolates of TcI and two of TcII. No polymorphic sites were found among sequences from 3 isolates of each TcIII and TcIV. Sequences from Non-Esmeraldo-like haplotype of CL Brener were identical to those of TcIII (Fig. 4).

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