Trypanosomatids regulate gene expression mainly at the post-transcriptional level through processing, exporting and stabilising mRNA and control of translation. In most eukaryotes, protein synthesis is regulated by phosphorylation of eukaryotic initiation factor 2 (eIF2) at serine 51. Phosphorylation halts overall translation by decreasing availability of initiator tRNAmet to form translating ribosomes. In trypanosomatids, the N-terminus of eIF2α is extended with threonine 169 the homologous phosphorylated residue. Here, we evaluated whether eIF2α phosphorylation varies during the Trypanosoma cruzi life cycle, the etiological agent of Chagas' disease. Total levels of eIF2α are diminished in infective and non-replicative trypomastigotes compared with proliferative forms from the intestine of the insect vector or amastigotes from mammalian cells, consistent with decreased protein synthesis reported in infective forms. eIF2α phosphorylation increases in proliferative intracellular forms prior to differentiation into trypomastigotes. Parasites overexpressing eIF2αT169A or with an endogenous CRISPR/Cas9-generated eIF2αT169A mutation were created and analysis revealed alterations to the proteome, largely unrelated to the presence of μORF in epimastigotes. eIF2αT169A mutant parasites produced fewer trypomastigotes with lower infectivity than wild type, with increased levels of sialylated mucins and oligomannose glycoproteins, and decreased galactofuranose epitopes and the surface protease GP63 on the cell surface. We conclude that eIF2α expression and phosphorylation levels affect proteins relevant for intracellular progression of T. cruzi.

• Klíčová slova
• Trypanosoma cruzi, differentiation, eIF2, phosphorylation, translation, virulence,
• MeSH
• buněčné linie MeSH
• Chagasova nemoc parazitologie   MeSH
• CRISPR-Cas systémy MeSH
• eukaryotický iniciační faktor 2 genetika   metabolismus   MeSH
• fosforylace MeSH
• lidé MeSH
• mutace MeSH
• nádorové buněčné linie MeSH
• parazitemie MeSH
• proteom metabolismus   MeSH
• proteosyntéza MeSH
• protozoální proteiny analýza   biosyntéza   metabolismus   MeSH
• regulace genové exprese MeSH
• stadia vývoje MeSH
• Trypanosoma cruzi růst a vývoj   metabolismus   patogenita   MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• eukaryotický iniciační faktor 2 MeSH
• proteom MeSH
• protozoální proteiny MeSH