Quantitative proteome and phosphoproteome analyses highlight the adherent population during Trypanosoma cruzi metacyclogenesis

Abstract

Trypanosoma cruzi metacyclogenesis is a natural process that occurs inside the triatomine vector and corresponds to the differentiation of non-infective epimastigotes into infective metacyclic trypomastigotes. The biochemical alterations necessary for the differentiation process have been widely studied with a focus on adhesion and nutritional stress. Here, using a mass spectrometry approach, a large-scale phospho(proteome) study was performed with the aim of understanding the metacyclogenesis processes in a quantitative manner. The results indicate that major modulations in the phospho(proteome) occur under nutritional stress and after 12 and 24 h of adhesion. Significant changes involve key cellular processes, such as translation, oxidative stress, and the metabolism of macromolecules, including proteins, lipids, and carbohydrates. Analysis of the signalling triggered by kinases and phosphatases from 7,336 identified phosphorylation sites demonstrates that 260 of these sites are modulated throughout the differentiation process, and some of these modulated proteins have previously been identified as drug targets in trypanosomiasis treatment. To the best of our knowledge, this study provides the first quantitative results highlighting the modulation of phosphorylation sites during metacyclogenesis and the greater coverage of the proteome to the parasite during this process. The data are available via ProteomeXchange with identifier number PXD006171.

Adenylate Cyclases of Trypanosoma brucei, Environmental Sensors and Controllers of Host Innate Immune Response

“…The presence of multiple putative phosphorylation sites that are conserved in the C-terminal region of the ACs (including the catalytic domain) suggests that this region may be involved in regulation of AC activity [ 88 ]. In T. cruzi , a quantitative proteomic and phosphoproteomic analysis confirmed that the C-terminal region of an AC isoform upregulated during metacyclogenesis [ 88 ] possesses 6 phosphorylation sites [ 89 ], which are partially conserved among the different T. brucei AC isoforms and may play a role in modulation of AC activity. A similar feature has been described for the kinase homology domain of NPR-A GC [ 90 ].…”

Study of VIPER and TATE in kinetoplastids and the evolution of tyrosine recombinase retrotransposons

“…However, we need to be cautious concluding TE activity in trypanosomatids based only on this type of data, since those reads could be only a transitory molecule resulting from global RNA expression. On the other hand, peptides from Gag-like protein were detected during the differentiation process from T. cruzi Dm28c noninfective epimastigotes into infective metacyclic trypomastigotes [68], a stronger sign of possible activity. In addition, we found potential encoding VIPER copies in some other species.…”

“…To detect VIPER activity in T. cruzi Dm28c, we analyzed the presence of RNA-seq reads and peptides from VIPER in results of published transcriptomic [67] and proteomic studies of this strain [68]. As most VIPER copies are annotated as hypothetical proteins, we retrieved all TriTrypDB gene IDs and UniProt IDs from VIPER copies and searched for them in the tables containing all genes and proteins identified in the studies using in-house Python code.…”

Redox Balance Keepers and Possible Cell Functions Managed by Redox Homeostasis in Trypanosoma cruzi

“…Schenkman and co-workers have recently developed the acetylome of T. cruzi, and showed that some cytosolic enzymes including antioxidant effectors were continuously acetylated and deacetylated; presumably as a control system for their enzymatic activity or protein-protein interaction (Moretti et al, 2018). Further, a large-scale phosphoproteome analysis study showed that antioxidant enzymes, TryR, TryS, and thiol-dependent reductase 1, are major targets of phosphorylation during the metacyclogenesis transition process (Marchini et al, 2011;Amorim et al, 2017). Thus, PTMs could have a key role in achieving a rapid regulation of antioxidants enzymatic activity as well as in stage-specific modulation of antioxidants expression.…”