Describing new players at Trypanosoma cruzi chromatin by locus-specific chromatin immunoprecipitation

• Grant number: 19/19834-5
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: February 01, 2020
• End date: January 31, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Julia Pinheiro Chagas da Cunha
• Grantee: Hellida Marina Costa Silva
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated research grant: 18/15553-9 - Going deeper into Trypanosoma cruzi chromatin regulation: identifying new players and quizzing its impact on a potential transcription control, AP.JP2

Abstract

At JP1, our group described many chromatin-associated proteins as well as more than 40 histone PTMs differentially expressed in replicate versus non-replicative life forms in Trypanosoma cruzi. Classical eu- and heterochromatin regions are presented at their nucleus and changes dramatically during differentiation from epimastigotes to metacyclics (E-M). Traditionally, these chromatin regions (and some histone PTMs) are associated to transcription regulation. These dataset, however, do not allow the association of a given protein (or a set of) to a specific genomic locus. That would, in turn, increase the comprehensive of chromatin biology possibly describing unpredicted functions of both protein and the genomic context. Recently, cutting-edge methodologies have been developed to recover proteins associated to a given locus. Thus, here we will evaluate the chromatin content of specific regions of T.cruzi genome by using dCas9-Flag (both in vitro and in vivo strategies) guided to target specific genomic regions (namely, TTS, TSS, telomeric regions, replication origins, SL promoter regions; 24S and small-subunit rRNA (RNA Pol I transcripts); tRNA and soRNA regions (RNA Pol III transcripts)). The comparison among the associated-chromatin content of each locus, together with the evaluation of (possible) changes during differentiation E-M (and cell cycle) may contribute for a better understanding of factors involved on the establishment and maintenance of gene silencing/activation as well as describe key protein components associated to differentiation/cell cycle. Finally, the role of 1-2 targets will be explored through CRISPR/Cas9 system for generation of knockouts (KO) parasites and tagged proteins.

Scientific publications

(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)

JERONIMO LIMA, ALEX RANIERI; DE SOUSA SILVA, HERBERT GUIMARAES; POUBEL, SALOE; ROSON, JULIANA NUNES; OLIVEIRA DE LIMA, LOYZE PAOLA; COSTA-SILVA, HELLIDA MARINA; GONCALVES, CAMILA SILVA; GALANTE, PEDRO A. F.; HOLETZ, FABIOLA; MOTTA, MARIA CRISTINA MACHADO M.; et al. Open chromatin analysis in Trypanosoma cruzi life forms highlights critical differences in genomic compartments and developmental regulation at tDNA loci. EPIGENETICS & CHROMATIN, v. 15, n. 1, p. 16-pg., 2022-06-01. (13/07467-1, 18/15553-9, 20/02708-4, 18/14432-3, 19/19834-5, 19/19690-3)

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, FEB 2022. (18/15553-9, 16/50050-2, 17/06104-3, 19/04483-2, 19/16033-1, 13/07467-1, 19/19834-5)