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An Insight into the proteome of Crithidia fasciculata choanomastigotes as a comparative approach to axenic growth, peanut lectin agglutination and differentiation of Leishmania spp. promastigotes.

Alcolea PJ, Alonso A, García-Tabares F, Toraño A, Larraga V - PLoS ONE (2014)

Bottom Line: A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein.The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation.The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Biology of Infections and Service of Proteomics and Genomics, Centro de Investigaciones Biológicas (Consejo Superior de Investigaciones Científicas), Madrid, Spain.

ABSTRACT
The life cycle of the trypanosomatid Crithidia fasciculata is monogenetic, as the unique hosts of these parasites are different species of culicids. The comparison of these non-pathogenic microorganisms evolutionary close to other species of trypanosomatids that develop digenetic life cycles and cause chronic severe sickness to millions of people worldwide is of outstanding interest. A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein. The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation. As L. major and L. infantum agglutinate with peanut lectin and non-agglutinating parasites are more infective, the agglutination properties were evaluated in C. fasciculata. The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated. As a difference with L. infantum, the non-agglutinating subpopulation over-expresses the whole machinery for maintenance of redox homeostasis and the translation factors eIF5a, EF1α and EF2, what suggests a relationship between the lack of agglutination and a differentiation process.

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Differentially expressed proteins involved in redox homeostasis and translation in C. fasciculata choanomastigotes.(A) The redox control system. Legend: proteins/protein variants in blue are constitutively expressed throughout the growth curve; proteins/protein variants in red are up-regulated at day 1 or 2 (logarithmic phase); proteins in purple are up-regulated in PNA- choanomastigotes. (B) Summary of differential abundance of translation factors and enzymes involved in redox homeostasis throughout the growth curve and in the PNA- subpopulation of choanomastigotes in stationary phase.
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pone-0113837-g005: Differentially expressed proteins involved in redox homeostasis and translation in C. fasciculata choanomastigotes.(A) The redox control system. Legend: proteins/protein variants in blue are constitutively expressed throughout the growth curve; proteins/protein variants in red are up-regulated at day 1 or 2 (logarithmic phase); proteins in purple are up-regulated in PNA- choanomastigotes. (B) Summary of differential abundance of translation factors and enzymes involved in redox homeostasis throughout the growth curve and in the PNA- subpopulation of choanomastigotes in stationary phase.

Mentions: The TryP of C. fasciculata previously characterized [40] (gi3851500; CfaCl_10_1430) is over-expressed in mid logarithmic phase choanomastigotes, as well as the thiol-dependent reductase 1 (TDR1) (Table 1, Fig. 5). This is an important difference with promastigotes of L. major, an ethiological agent of cutaneous leishmaniasis in the Old World, as TryP is constitutively expressed in all the stages of its life cycle [41], [42]. It is also known that L. donovani amastigotes up-regulate the TryP with respect to promastigotes [27]. The catalase is absent in pathogenic trypanosomatids [43] but not in C. fasciculata. In fact, variants matching with the annotations CfaCl_30_0050 have been identified in several spots (Tables 1 and 3). For this reason, hydrogen peroxide removal in C. fasciculata is not necessarily dependent on trypanothione-linked peroxidases, as a difference with the pathogenic trypanosomatids. Like the TryP, this protein is more abundant at mid logarithmic phase. These data suggest higher levels of oxidative stress counteracted with TDR1, TryP and catalase up-regulation in Crithidia spp. choanomastigotes than in Leishmania spp. promastigotes at mid logarithmic phase, possibly due to the greater growth rate observed in the former (Fig. 1). In fact, the stationary phase is reached about 5–7 days in a typical growth curve of Leishmania spp. (e.g., [24]). The up-regulation of TDR1 suggests that the glutathione-ascorbate cycle is also participating in counteracting oxidative stress.


An Insight into the proteome of Crithidia fasciculata choanomastigotes as a comparative approach to axenic growth, peanut lectin agglutination and differentiation of Leishmania spp. promastigotes.

Alcolea PJ, Alonso A, García-Tabares F, Toraño A, Larraga V - PLoS ONE (2014)

Differentially expressed proteins involved in redox homeostasis and translation in C. fasciculata choanomastigotes.(A) The redox control system. Legend: proteins/protein variants in blue are constitutively expressed throughout the growth curve; proteins/protein variants in red are up-regulated at day 1 or 2 (logarithmic phase); proteins in purple are up-regulated in PNA- choanomastigotes. (B) Summary of differential abundance of translation factors and enzymes involved in redox homeostasis throughout the growth curve and in the PNA- subpopulation of choanomastigotes in stationary phase.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263474&req=5

pone-0113837-g005: Differentially expressed proteins involved in redox homeostasis and translation in C. fasciculata choanomastigotes.(A) The redox control system. Legend: proteins/protein variants in blue are constitutively expressed throughout the growth curve; proteins/protein variants in red are up-regulated at day 1 or 2 (logarithmic phase); proteins in purple are up-regulated in PNA- choanomastigotes. (B) Summary of differential abundance of translation factors and enzymes involved in redox homeostasis throughout the growth curve and in the PNA- subpopulation of choanomastigotes in stationary phase.
Mentions: The TryP of C. fasciculata previously characterized [40] (gi3851500; CfaCl_10_1430) is over-expressed in mid logarithmic phase choanomastigotes, as well as the thiol-dependent reductase 1 (TDR1) (Table 1, Fig. 5). This is an important difference with promastigotes of L. major, an ethiological agent of cutaneous leishmaniasis in the Old World, as TryP is constitutively expressed in all the stages of its life cycle [41], [42]. It is also known that L. donovani amastigotes up-regulate the TryP with respect to promastigotes [27]. The catalase is absent in pathogenic trypanosomatids [43] but not in C. fasciculata. In fact, variants matching with the annotations CfaCl_30_0050 have been identified in several spots (Tables 1 and 3). For this reason, hydrogen peroxide removal in C. fasciculata is not necessarily dependent on trypanothione-linked peroxidases, as a difference with the pathogenic trypanosomatids. Like the TryP, this protein is more abundant at mid logarithmic phase. These data suggest higher levels of oxidative stress counteracted with TDR1, TryP and catalase up-regulation in Crithidia spp. choanomastigotes than in Leishmania spp. promastigotes at mid logarithmic phase, possibly due to the greater growth rate observed in the former (Fig. 1). In fact, the stationary phase is reached about 5–7 days in a typical growth curve of Leishmania spp. (e.g., [24]). The up-regulation of TDR1 suggests that the glutathione-ascorbate cycle is also participating in counteracting oxidative stress.

Bottom Line: A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein.The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation.The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Biology of Infections and Service of Proteomics and Genomics, Centro de Investigaciones Biológicas (Consejo Superior de Investigaciones Científicas), Madrid, Spain.

ABSTRACT
The life cycle of the trypanosomatid Crithidia fasciculata is monogenetic, as the unique hosts of these parasites are different species of culicids. The comparison of these non-pathogenic microorganisms evolutionary close to other species of trypanosomatids that develop digenetic life cycles and cause chronic severe sickness to millions of people worldwide is of outstanding interest. A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein. The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation. As L. major and L. infantum agglutinate with peanut lectin and non-agglutinating parasites are more infective, the agglutination properties were evaluated in C. fasciculata. The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated. As a difference with L. infantum, the non-agglutinating subpopulation over-expresses the whole machinery for maintenance of redox homeostasis and the translation factors eIF5a, EF1α and EF2, what suggests a relationship between the lack of agglutination and a differentiation process.

Show MeSH
Related in: MedlinePlus