Improvements on the quantitative analysis of Trypanosoma cruzi histone post translational modifications: Study of changes in epigenetic marks through the parasite's metacyclogenesis and life cycle

Autor(es)

de Lima, Loyze P. ^{[1, 2]} ; Poubel, Saloe Bispo ^{[1, 2, 3]} ; Yuan, Zuo-Fei ^[4] ; Roson, Juliana Nunes ^{[1, 2]} ; de Luna Vitorino, Francisca Nathalia ^{[1, 2]} ; Holetz, Fabiola Barbieri ^[3] ; Garcia, Benjamin A. ^[4] ; Chagas da Cunha, Julia Pinheiro ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Inst Butantan, Lab Ciclo Celular, Sao Paulo, SP - Brazil

^[2] Inst Butantan, Ctr Toxins Immune Response & Cell Signaling CeTIC, Sao Paulo, SP - Brazil

^[3] Fiocruz MS, Inst Carlos Chagas, Rua Algacyr Munhoz Mader, 3775 CIC, BR-81350010 Curitiba, PR - Brazil

^[4] Univ Penn, Perelman Sch Med, Dept Biochem & Biophys, Epigenet Inst, Philadelphia, PA 19104 - USA

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF PROTEOMICS; v. 225, APR 15 2020.
• Citações Web of Science: 0

Resumo

Trypanosome histone N-terminal sequences are very divergent from the other eukaryotes, although they are still decorated by post-translational modifications (PTMs). Here, we used a highly robust workflow to analyze histone PTMs in the parasite Trypanosoma cruzi using mass spectrometry-based (MS-based) data-independent acquisition (DIA). We adapted the workflow for the analysis of the parasite's histone sequences by modifying the software EpiProfile 2.0, improving peptide and PTM quantification accuracy. This workflow could now be applied to the study of 141 T. cruzi modified histone peptides, which we used to investigate the dynamics of histone PTMs along the metacyclogenesis and the life cycle of T. cruzi. Global levels of histone acetylation and methylation fluctuates along metacyclogenesis, however most critical differences were observed between parasite life forms. More than 66 histone PTM changes were detected. Strikingly, the histone PTM pattern of metacyclic trypomastigotes is more similar to epimastigotes than to cellular trypomastigotes. Finally, we highlighted changes at the H4 N-terminus and at H3K76 discussing their impact on the trypanosome biology. Altogether, we have optimized a workflow easily applicable to the analysis of histone PTMs in T. cruzi and generated a dataset that may shed lights on the role of chromatin modifications in this parasite. Significance: Trypanosomes are unicellular parasites that have divergent histone sequences, no chromosome condensation and a peculiar genome/gene regulation. Genes are transcribed from divergent polycistronic regions and post-transcriptional gene regulation play major role on the establishment of transcripts and protein levels. In this regard, the fact that their histones are decorated with multiple PTMs raises interesting questions about their role. Besides, this digenetic organism must adapt to different environments changing its metabolism accordingly. As metabolism and epigenetics are closely related, the study of histone PTMs in trypanosomes may enlighten this strikingly, and not yet fully understood, interplay. From a biomedical perspective, the comprehensive study of molecular mechanisms associated to the metacyclogenesis process is essential to create better strategies for controlling Chagas disease. (AU)

• Processo FAPESP: 18/21785-0 - Análise da interação Orc1/Cdc6-DNA durante o ciclo de vida de Trypanosoma cruzi
• Beneficiário: Loyze Paola Oliveira de Lima
• Linha de fomento: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/06104-3 - Análise funcional da histona H2B variante de Trypanosoma cruzi
• Beneficiário: Julia Pinheiro Chagas da Cunha
• Linha de fomento: Auxílio à Pesquisa - Regular
• Processo FAPESP: 18/14432-3 - Uma rede para uma biologia integrativa em doenças negligenciadas: conectando a epigenética, o metabolismo e a biologia celular em tripanossomatídeos patogênicos
• Beneficiário: Ariel Mariano Silber
• Linha de fomento: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/15553-9 - Indo a fundo na regulação da cromatina de Trypanosoma cruzi: identificação de novas moléculas e questionando seu possível impacto no controle da transcrição
• Beneficiário: Julia Pinheiro Chagas da Cunha
• Linha de fomento: Auxílio à Pesquisa - Jovens Pesquisadores - Fase 2
• Processo FAPESP: 13/07467-1 - CeTICS - Centro de Toxinas, Imuno-Resposta e Sinalização Celular
• Beneficiário: Hugo Aguirre Armelin
• Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 17/18344-9 - Proteômica quantitativa da cromatina frente ao tratamento com FGF2: análise da regulação transcricional e associação de cdc42
• Beneficiário: Francisca Nathália de Luna Vitorino
• Linha de fomento: Bolsas no Brasil - Doutorado Direto