RNA-Seq reveals that overexpression of TcUBP1 switches the gene expression pattern toward that of the infective form of Trypanosoma cruzi

Transcriptomic analysis of N-terminal mutated Trypanosoma cruzi UBP1 knockdown underlines the importance of this RNA-binding protein in parasite development

“…Our previous findings revealed that overexpression of TcUBP1 in insect-dwelling epimastigotes enhanced the abundance of TcS transcripts and altered their subcellular localization to a perinuclear region [36,46]. These observations were further supported by a shift toward mRNA expression characteristic of infective trypomastigotes [16,36]. These findings agree with previous reports indicating a three-fold increase in TcUBP1 protein levels during parasite differentiation into metacyclic trypomastigotes [47].…”

“…These results suggest that the mRNA abundances of members of the TcS superfamily are directly influenced by TcUBP1 levels, both increasing their expression when UBP1 protein levels are high and decreasing their expression when UBP1 protein levels are low. By contrast, the main cluster among the downregulated genes previously reported in the OE parasite transcriptome was that coding for ribosomal proteins [16]. Within this functional category, we found 15 genes whose abundance varied in a coordinated way in both OE and KD conditions (|log2 fold change| >0.45, FDR-adjusted p value < 5%), Fig 8B and Table 4).…”

“…establishing a replicative or infective profile (ribosomal proteins, MASP, MUCI/II and TcS), respond directly and proportionally to TcUBP1 protein levels, indicating that this RBP plays a pivotal role in either triggering or not the differentiation process. The most abundant cluster among the upregulated genes in the OE transcriptome was that coding for trypomastigote cell-surface glycoproteins [16]. Within the TcS superfamily group, we identified 42 significant robust genes (Table 3), whose abundance in both OE and KD 8A).…”

“…RNA-binding proteins (RBPs) that are bound to an mRNA determine its fate within the cell, affecting translation [5][6][7], nuclear export [8], mRNA stability [9], and mRNA processing [10]. RBPs can act as critical trans-acting factors at each of these regulatory points and mediate parasite differentiation in both T. cruzi and T. brucei [11][12][13][14][15][16].…”

RNA-seq data exploration after trypanosome RNA-binding protein UBP1 expression is altered by CRISPR-Cas9 gene editing and overexpression

“…Remarkably, UBP1 gene is encoded in a single stable dicistronic unit and its protein product regulates the abundance of several genes that have U-rich elements in Trypanosoma cruzi [ 1 , 2 ]. We have already shown that when UBP1 is overexpressed in insect-dwelling epimastigotes, the microorganism shows an expression pattern that looks like the infective form [3] . To further investigate the regulatory role of this protein, we used the CRISPR-Cas9 technology to generate a population of parasites that did not express this protein.…”