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Memórias do Instituto Oswaldo Cruz

versão impressa ISSN 0074-0276versão On-line ISSN 1678-8060

Resumo

SANTOS, Cyndia Mara Bezerra dos et al. Trypanosoma cruzi transcriptome during axenic epimastigote growth curve. Mem. Inst. Oswaldo Cruz [online]. 2018, vol.113, n.5, e170404.  Epub 09-Abr-2018. ISSN 0074-0276.  http://dx.doi.org/10.1590/0074-02760170404.

BACKGROUND

Trypanosoma cruzi is an important protozoan parasite and the causative agent of Chagas disease. A critical step in understanding T. cruzi biology is the study of cellular and molecular features exhibited during its growth curve.

OBJECTIVES

We aimed to acquire a global view of the gene expression profile of T. cruzi during epimastigote growth.

METHODS

RNA-Seq analysis of total and polysomal/granular RNA fractions was performed along the 10 days T. cruzi epimastigote growth curve in vitro, in addition to cell viability and cell cycle analyses. We also analysed the polysome profile and investigated the presence of granular RNA by FISH and western blotting.

FINDINGS

We identified 1082 differentially expressed genes (DEGs), of which 220 were modulated in both fractions. According to the modulation pattern, DEGs were grouped into 12 clusters and showed enrichment of important gene ontology (GO) terms. Moreover, we showed that by the sixth day of the growth curve, polysomal content declined greatly and the RNA granules content appeared to increase, suggesting that a portion of mRNAs isolated from the sucrose gradient during late growth stages was associated with RNA granules and not only polyribosomes. Furthermore, we discuss several modulated genes possibly involved in T. cruzi growth, mainly during the stationary phase, such as genes related to cell cycle, pathogenesis, metabolic processes and RNA-binding proteins.

Palavras-chave : Chagas disease; growth curve; transcriptomics; RNA-Seq; polysomes; RNA granules.

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