EIF2 alpha phosphorylation is regulated in intracellular amastigotes for the generation of infectiveTrypanosoma cruzitrypomastigote forms

Autor(es)

Castro Machado, Fabricio ^[1] ; Bittencourt-Cunha, Paula ^[1] ; Malvezzi, Amaranta Muniz ^[1] ; Arico, Mirella ^[1] ; Radio, Santiago ^{[2, 3]} ; Smircich, Pablo ^{[2, 3]} ; Zoltner, Martin ^[4] ; Field, Mark C. ^{[5, 6]} ; Schenkman, Sergio ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Dept Microbiol Imunol & Parasitol, Escola Paulista Med, R Pedro Toledo 669 L6A, BR-04039032 Sao Paulo, SP - Brazil

^[2] Minist Educ & Cultura, Dept Genom, Inst Invest Biol Clemente Estable, Montevideo - Uruguay

^[3] Univ Republica, Fac Ciencias, Lab Mol Interact, Montevideo - Uruguay

^[4] Charles Univ Prague, Ctr Res Pathogen & Virulence Parasites, Drug Discovery & Evaluat, Prague - Czech Republic

^[5] Univ Dundee, Div Biol Chem & Drug Discovery, Dundee - Scotland

^[6] Czech Acad Sci, Inst Parasitol, Prague - Czech Republic

• Tipo de documento: Artigo Científico
• Fonte: Cellular Microbiology; v. 22, n. 11 JUL 2020.
• Citações Web of Science: 0

Resumo

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 tRNA(met)to form translating ribosomes. In trypanosomatids, theN-terminus of eIF2 alpha is extended with threonine 169 the homologous phosphorylated residue. Here, we evaluated whether eIF2 alpha phosphorylation varies during theTrypanosoma cruzilife cycle, the etiological agent of Chagas' disease. Total levels of eIF2 alpha 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. eIF2 alpha phosphorylation increases in proliferative intracellular forms prior to differentiation into trypomastigotes. Parasites overexpressing eIF2 alpha(T169A)or with an endogenous CRISPR/Cas9-generated eIF2 alpha(T169A)mutation were created and analysis revealed alterations to the proteome, largely unrelated to the presence of mu ORF in epimastigotes. eIF2 alpha(T169A)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 eIF2 alpha expression and phosphorylation levels affect proteins relevant for intracellular progression ofT. cruzi. (AU)

• Processo FAPESP: 14/01577-2 - O papel da fosforilação de eIF2 e de sirtuinas na multiplicação e diferenciação intracelular de Trypanosoma cruzi.
• Beneficiário: Fabrício Castro Machado
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 17/02496-4 - Papel da acetilação de proteínas e sirtuínas no processo de infecção pelo T. cruzi
• Beneficiário: Amaranta Muniz Malvezzi
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado