Unveiling the Trypanosoma cruzi Nuclear Proteome

Júnior, Agenor de Castro Moreira dos Santos; Kalume, Dário Eluan; Camargo, Ricardo; Gómez‐Mendoza, Diana Paola; Corrêa, José R.; Charneau, Sébastien; Sousa, Marcelo Valle de; Lima, Beatriz Dolabela de; Ricart, Carlos André Ornelas

doi:10.1371/journal.pone.0138667

Cited by 19 publications

(13 citation statements)

References 39 publications

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“…We found that RHS proteins are located in the nucleus of epimastigotes and amastigotes of T. cruzi. This is in agreement with previous work [54] that identified the presence of 74 RHS proteins with apparent molecular masses of 12 to 111 kDa in the nuclear proteome of T. cruzi epimastigotes [54]. These data were corroborated by Western blot analysis, in which we identified RHS proteins from 29 to 118 kDa in CLB.…”

Section: The Role Of Rhs Proteins In T Cruzisupporting

confidence: 93%

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Genomic Organization and Generation of Genetic Variability in the RHS (Retrotransposon Hot Spot) Protein Multigene Family in Trypanosoma cruzi

Bernardo

Souza

Costa-Martins

et al. 2020

Genes

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Retrotransposon Hot Spot (RHS) is the most abundant gene family in Trypanosoma cruzi, with unknown function in this parasite. The aim of this work was to shed light on the organization and expression of RHS in T. cruzi. The diversity of the RHS protein family in T. cruzi was demonstrated by phylogenetic and recombination analyses. Transcribed sequences carrying the RHS domain were classified into ten distinct groups of monophyletic origin. We identified numerous recombination events among the RHS and traced the origins of the donors and target sequences. The transcribed RHS genes have a mosaic structure that may contain fragments of different RHS inserted in the target sequence. About 30% of RHS sequences are located in the subtelomere, a region very susceptible to recombination. The evolution of the RHS family has been marked by many events, including gene duplication by unequal mitotic crossing-over, homologous, as well as ectopic recombination, and gene conversion. The expression of RHS was analyzed by immunofluorescence and immunoblotting using anti-RHS antibodies. RHS proteins are evenly distributed in the nuclear region of T. cruzi replicative forms (amastigote and epimastigote), suggesting that they could be involved in the control of the chromatin structure and gene expression, as has been proposed for T. brucei.

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Section: The Role Of Rhs Proteins In T Cruzisupporting

confidence: 93%

“…This region is conserved among RHS of some T. cruzi strains (Dm28c, Sylvio X10/1, Y, Bug2148, Tulahuen, TCC) and T. cruzi marinkellei. The location of RHS (TcCLB.511055.20) in the nucleus has been experimentally demonstrated in the nuclear subproteome of clone CLB [54].…”

Section: Expression and Subcellular Localization Of Rhs In T Cruzimentioning

confidence: 94%

Genomic Organization and Generation of Genetic Variability in the RHS (Retrotransposon Hot Spot) Protein Multigene Family in Trypanosoma cruzi

Bernardo

Souza

Costa-Martins

et al. 2020

Genes

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“…Multiple copies of RHS genes have been annotated in all African trypanosomes ( T. brucei, T. congolense, T. vivax ) and in other members of the genus Trypanosoma , such as T. grayi, T. theileri and T. cruzi. T. cruzi RHS have been identified in the nuclear proteome and shown to be upregulated after heat shock, but nothing is known about their function (97,98). Leishmania spp., despite belonging to the trypanosomatids, have no RHS genes.…”

Section: Discussionmentioning

confidence: 99%

Unexpected diversity in eukaryotic transcription revealed by the retrotransposon hotspot family of Trypanosoma brucei

Florini

Naguleswaran

Gharib

et al. 2018

Nucleic Acids Research

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The path from DNA to RNA to protein in eukaryotes is guided by a series of factors linking transcription, mRNA export and translation. Many of these are conserved from yeast to humans. Trypanosomatids, which diverged early in the eukaryotic lineage, exhibit unusual features such as polycistronic transcription and trans-splicing of all messenger RNAs. They possess basal transcription factors, but lack recognisable orthologues of many factors required for transcription elongation and mRNA export. We show that retrotransposon hotspot (RHS) proteins fulfil some of these functions and that their depletion globally impairs nascent RNA synthesis by RNA polymerase II. Three sub-families are part of a coordinated process in which RHS6 is most closely associated with chromatin, RHS4 is part of the Pol II complex and RHS2 connects transcription with the translation machinery. In summary, our results show that the components of eukaryotic transcription are far from being universal, and reveal unsuspected plasticity in the course of evolution.

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“…serpens (strain 9T) was grown in brain heart infusion broth (BHI) medium (Acumedia, Lansing, Michigan) at 27 °C. Parasite cells (5 x 10 8 ) collected in logarithmic growth phase were fractionated using a protocol based on a previously described report [ 25 ]. Parasites were centrifuged for 10 min at 5,000 g then washed three times in phosphate buffered saline (PBS).…”

Section: Methodsmentioning

confidence: 99%

Proteome analysis of Phytomonas serpens, a phytoparasite of medical interest

et al. 2018

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The protozoan Phytomonas serpens (class Kinetoplastea) is an important phytoparasite that has gained medical importance due to its similarities to Trypanosoma cruzi, the etiological agent of Chagas disease. The present work describes the first proteome analysis of P. serpens. The parasite was separated into cytosolic and high density organelle fractions, which, together with total cell extract, were subjected to LC-MS/MS analyses. Protein identification was conducted using a comprehensive database composed of genome sequences of other related kinetoplastids. A total of 1,540 protein groups were identified among the three sample fractions. Sequences from Phytomonas sp. in the database allowed the highest number of identifications, with T. cruzi and T. brucei the human pathogens providing the greatest contribution to the identifications. Based on the proteomics data obtained, we proposed a central metabolic map of P. serpens, which includes all enzymes of the citric acid cycle. Data also revealed a new range of proteins possibly responsible for immunological cross-reactivity between P. serpens and T. cruzi.

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Unveiling the Trypanosoma cruzi Nuclear Proteome

Cited by 19 publications

References 39 publications

Genomic Organization and Generation of Genetic Variability in the RHS (Retrotransposon Hot Spot) Protein Multigene Family in Trypanosoma cruzi

Genomic Organization and Generation of Genetic Variability in the RHS (Retrotransposon Hot Spot) Protein Multigene Family in Trypanosoma cruzi

Unexpected diversity in eukaryotic transcription revealed by the retrotransposon hotspot family of Trypanosoma brucei

Proteome analysis of Phytomonas serpens, a phytoparasite of medical interest

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