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Functional analysis of Trypanosoma Cruzi histone H2B variant


Trypanosoma cruzi is a unicellular eukaryote, a causative agent of Chagas disease that alternates between replicative and non-infective forms (epimastigotes and amastigotes) and non-replicative and infective forms (cellular and blood trypomastigotes). Differentiation between these forms is accompanied by differences in overall transcript levels and structural changes in chromatin. Several studies indicate that trypanosome chromatin contains several epigenetic alterations, including deposition of histone variants, enriched in putative transcription start sites. We recently verified that histone H2Bv of T.cruzi is enriched at chromatin from non-replicative forms, suggesting that it may play important roles in the phenotypic differences (gene expression and chromatin structure) observed between life forms of T. cruzi. Thus, in this work, we intend to study the role of histone H2Bv in this parasite. To do this, we will: I. Identify the histone variant interactors' by co-immunoprecipitation and pulldown assays; II. identify the genomic regions in which this protein is associated by chromatin immunoprecipitation assays (ChIP); III. analyze if H2Bv is preferentially associated to euchromatin or heterochromatin by electron microscopy and partial digestion with MNase; IV. generate parasite knockouts for H2Bv and evaluate the phenotype during life cycle; V. Evaluate the phenotype of epimastigote forms and mammalian cells that overexpress H2Bv. (AU)

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(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DE LIMA, LOYZE P.; POUBEL, SALOE BISPO; YUAN, ZUO-FEI; ROSON, JULIANA NUNES; DE LUNA VITORINO, FRANCISCA NATHALIA; HOLETZ, FABIOLA BARBIERI; GARCIA, BENJAMIN A.; CHAGAS DA CUNHA, JULIA PINHEIRO. 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. JOURNAL OF PROTEOMICS, v. 225, . (18/21785-0, 17/06104-3, 18/14432-3, 18/15553-9, 13/07467-1, 17/18344-9)
ROSON, JULIANA NUNES; VITARELLI, MARCELA DE OLIVEIRA J.; COSTA-SILVA, HELLIDA MARINA; PEREIRA, KAMILLE SCHMITT; PIRES, DAVID DA SILVA; LOPES, LETICIA DE SOUSA; CORDEIRO, BARBARA; KRAUS, AMELIE; CRUZ, KARIN NAVARRO TOZZI; CALDERANO, SIMONE GUEDES; et al. 2B.V demarcates divergent strand-switch regions, some tDNA loci, and genome compartments in Trypanosoma cruzi and affects parasite differentiation and host cell invasio. PLOS PATHOGENS, v. 18, n. 2, . (18/15553-9, 16/50050-2, 17/06104-3, 19/04483-2, 19/16033-1, 13/07467-1, 19/19834-5)

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