Limits...
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.

Show MeSH

Related in: MedlinePlus

Alignment of predicted amino acid sequences from entire cruzipain of T. cruzi (TcI, TcII, TcIII, TcVI and Tcbat) and homologues from T. cruzi-like (T. c. marinkellei and T. dionisii), T. rangeli and T. b. brucei.Pre, pro, catalytic domain and C-terminal extension amino acid sequences of cruzipain genes from T. cruzi Sylvio X10.6 and G (TcI), TCC1994 (Tcbat), Y and Esmeraldo cl3 (TcII), M6241 cl6 (TcIII), CL Brener (TcVI) Non-Esmeraldo-like (TcIII) and Esmeraldo-like (TcII) haplotypes and homologues from T. c. marinkellei (344), T. dionisii (211), T. rangeli (LDG and AM80) and T. b. brucei (TREU 927). The CATL family signatures of pro-domain motifs ERFININ (ERFN) and GNFD (GTFD) are indicated in bold and underlined, the subsites S1, S2 and S2′ are in bold, and the conserved Trp181 are indicated by (*).The glutamine [Q] of the oxyanion hole, cysteine [C], histidine [H] and asparagine [N] of catalytic triad in the catalytic domain, and 8 cysteines in the C-terminal extension are indicated by arrow heads.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3369871&req=5

pone-0038385-g001: Alignment of predicted amino acid sequences from entire cruzipain of T. cruzi (TcI, TcII, TcIII, TcVI and Tcbat) and homologues from T. cruzi-like (T. c. marinkellei and T. dionisii), T. rangeli and T. b. brucei.Pre, pro, catalytic domain and C-terminal extension amino acid sequences of cruzipain genes from T. cruzi Sylvio X10.6 and G (TcI), TCC1994 (Tcbat), Y and Esmeraldo cl3 (TcII), M6241 cl6 (TcIII), CL Brener (TcVI) Non-Esmeraldo-like (TcIII) and Esmeraldo-like (TcII) haplotypes and homologues from T. c. marinkellei (344), T. dionisii (211), T. rangeli (LDG and AM80) and T. b. brucei (TREU 927). The CATL family signatures of pro-domain motifs ERFININ (ERFN) and GNFD (GTFD) are indicated in bold and underlined, the subsites S1, S2 and S2′ are in bold, and the conserved Trp181 are indicated by (*).The glutamine [Q] of the oxyanion hole, cysteine [C], histidine [H] and asparagine [N] of catalytic triad in the catalytic domain, and 8 cysteines in the C-terminal extension are indicated by arrow heads.

Mentions: Overall identities were high in the N-terminal region, either in pre- and pro-domains (∼94 and 91%, respectively) and catalytic domains (∼90%), and most variable in the C-terminal regions (∼85%) (Fig. 1). As typically found in peptidases of Clan CA, which are targeted to intracellular compartments and secreted, all cruzipain and homologous genes have a signal peptide at their N-terminal region, as well as the catalytic triad of cysteine, histidine and asparagine residues (Cys25, His159 and Asn179) and the highly conserved Trp181. Important sites for autocatalytic cleavage, the motifs ERFNIN-like and GNFD-like of pro-domains are conserved in all trypanosomes. The clan CA is characterized by having substrate specificity defined by the S2 pocket. In cruzipain genes from all T. cruzi DTUs and homologues from T. c. marinkellei, the S2 subsites are conserved whereas in T. dionisii, T. rangeli and T. b. brucei divergent amino acids were found in these regions, suggesting differences in substrate specificities (Fig. 1).


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)

Alignment of predicted amino acid sequences from entire cruzipain of T. cruzi (TcI, TcII, TcIII, TcVI and Tcbat) and homologues from T. cruzi-like (T. c. marinkellei and T. dionisii), T. rangeli and T. b. brucei.Pre, pro, catalytic domain and C-terminal extension amino acid sequences of cruzipain genes from T. cruzi Sylvio X10.6 and G (TcI), TCC1994 (Tcbat), Y and Esmeraldo cl3 (TcII), M6241 cl6 (TcIII), CL Brener (TcVI) Non-Esmeraldo-like (TcIII) and Esmeraldo-like (TcII) haplotypes and homologues from T. c. marinkellei (344), T. dionisii (211), T. rangeli (LDG and AM80) and T. b. brucei (TREU 927). The CATL family signatures of pro-domain motifs ERFININ (ERFN) and GNFD (GTFD) are indicated in bold and underlined, the subsites S1, S2 and S2′ are in bold, and the conserved Trp181 are indicated by (*).The glutamine [Q] of the oxyanion hole, cysteine [C], histidine [H] and asparagine [N] of catalytic triad in the catalytic domain, and 8 cysteines in the C-terminal extension are indicated by arrow heads.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0038385-g001: Alignment of predicted amino acid sequences from entire cruzipain of T. cruzi (TcI, TcII, TcIII, TcVI and Tcbat) and homologues from T. cruzi-like (T. c. marinkellei and T. dionisii), T. rangeli and T. b. brucei.Pre, pro, catalytic domain and C-terminal extension amino acid sequences of cruzipain genes from T. cruzi Sylvio X10.6 and G (TcI), TCC1994 (Tcbat), Y and Esmeraldo cl3 (TcII), M6241 cl6 (TcIII), CL Brener (TcVI) Non-Esmeraldo-like (TcIII) and Esmeraldo-like (TcII) haplotypes and homologues from T. c. marinkellei (344), T. dionisii (211), T. rangeli (LDG and AM80) and T. b. brucei (TREU 927). The CATL family signatures of pro-domain motifs ERFININ (ERFN) and GNFD (GTFD) are indicated in bold and underlined, the subsites S1, S2 and S2′ are in bold, and the conserved Trp181 are indicated by (*).The glutamine [Q] of the oxyanion hole, cysteine [C], histidine [H] and asparagine [N] of catalytic triad in the catalytic domain, and 8 cysteines in the C-terminal extension are indicated by arrow heads.
Mentions: Overall identities were high in the N-terminal region, either in pre- and pro-domains (∼94 and 91%, respectively) and catalytic domains (∼90%), and most variable in the C-terminal regions (∼85%) (Fig. 1). As typically found in peptidases of Clan CA, which are targeted to intracellular compartments and secreted, all cruzipain and homologous genes have a signal peptide at their N-terminal region, as well as the catalytic triad of cysteine, histidine and asparagine residues (Cys25, His159 and Asn179) and the highly conserved Trp181. Important sites for autocatalytic cleavage, the motifs ERFNIN-like and GNFD-like of pro-domains are conserved in all trypanosomes. The clan CA is characterized by having substrate specificity defined by the S2 pocket. In cruzipain genes from all T. cruzi DTUs and homologues from T. c. marinkellei, the S2 subsites are conserved whereas in T. dionisii, T. rangeli and T. b. brucei divergent amino acids were found in these regions, suggesting differences in substrate specificities (Fig. 1).

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