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Knocking out histidine ammonia-lyase by using CRISPR-Cas9 abolishes histidine role in the bioenergetics and the life cycle of Trypanosoma cruzi

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Author(s):
Nascimento, Janaina de Freitas ; Barison, Maria Julia ; Montanaro, Gabriela Torres ; Marchese, Leticia ; Souza, Rodolpho Ornitz Oliveira ; Silva, Leticia Sophia ; Guarnieri, Alessandra Aparecida ; Silber, Ariel Mariano
Total Authors: 8
Document type: Journal article
Source: MICROBIAL CELL; v. 12, n. 1, p. 16-pg., 2025-01-01.
Abstract

Trypanosoma cruzi, the causing agent of Chagas disease, is the only known trypanosomatid pathogenic to humans having a complete histidine to glutamate pathway, which involves a series of four enzymatic reactions that convert histidine into downstream metabolites, including urocanate, 4-imidazolone-5-propionate, N-formimino-L-glutamate and Lglutamate. Recent studies have highlighted the importance of this pathway in ATP production, redox balance, and the maintenance of cellular homeostasis in T. cruzi. In this work, we focus on the first step of the histidine degradation pathway, which is performed by the enzyme histidine ammonia lyase. Here we determined the kinetic and biochemical parameters of the T. cruzi histidine ammonia-lyase. By generating null mutants of this enzyme using CRISPR-Cas9 we observed that disruption of the first step of the histidine degradation pathway completely abolishes the capability of this parasite to metabolise histidine, compromising the use of this amino acid as an energy and carbon source. Additionally, we showed that the knockout of the histidine ammonia lyase affects metacyclogenesis when histidine is the only metabolizable source and diminishes trypomastigote infection in vitro. (AU)

FAPESP's process: 21/12938-0 - Amino acid metabolism in Trypanosoma cruzi: a toolbox to survive in hostile environments
Grantee:Ariel Mariano Silber
Support Opportunities: Research Projects - Thematic Grants