Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/16136
Title: A new antimicrobial protein from the anterior midgut of Triatoma infestans mediates Trypanosoma cruzi establishment by controlling the microbiota
Authors: Buarque, Diego de Souza
Gomes, Cícera Maria
Araújo, Ricardo Nascimento
Pereira, Marcos Horacio
Ferreira, Roberta Carvalho
Guarneri, Alessandra Aparecida
Tanaka, Aparecida Sadae
Affilliation: Universidade Federal de Sao Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. Sao Paulo, SP, Brasil
Universidade Federal de Sao Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. Sao Paulo, SP, Brasil
Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil
Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Belo Horizonte, MG, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Belo Horizonte, MG, Brasil
Universidade Federal de Sao Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. Sao Paulo, SP, Brasil
Abstract: The Reduviid Triatoma infestans is a vector for the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. The parasite must address the defense molecules and microbiota that colonize the anterior midgut of T. infestans. To obtain insight into T. cruzi - microbiota interactions in triatomine insects, we characterized a new antimicrobial product from the anterior midgut of T. infestans (TiAP) that may be involved in these relationships. The TiAP DNA fragment was cloned and expressed in a bacterial system, and the effect of the protein on bacteria and T. cruzi was evaluated by RNAi, qPCR and antimicrobial experiments. The number of T. cruzi in T. infestans anterior midguts was significantly lower in TiAP knockdown insects than in unsilenced groups. We also verified that the amount of bacteria in silenced T. infestans is approximately 600-fold higher than in unsilenced insects by qPCR. The 327-bp cDNA fragment that encodes mature TiAP was cloned into the pET-14b vector and expressed fused to a His-tag in Escherichia coli C43. The recombinant protein (rTiAP) was purified using an Ni-NTA column, followed by a HiTrap SP column. According to a trypanocidal assay, rTiAP did not interfere with the viability of T. cruzi trypomastigotes. Moreover, in antimicrobial experiments using E. coli and Micrococcus luteus, the protein was only bacteriostatic for Gram-negative bacteria. The data indicate that infection by T. cruzi increases the expression of TiAP to modulate the microbiota. The inhibition of microbiota growth by TiAP is important for parasite establishment in the T. infestans anterior midgut.
Keywords: Antimicrobial properties
Bacteria
Midgut
Parasite
Triatoma infestans
Trypanosoma cruzi
keywords: Propriedades antimicrobianas
Issue Date: 2016
Publisher: Elsevier B.V.
Citation: BUARQUE, Diego de Souza et al. A new antimicrobial protein from the anterior midgut of Triatoma infestans mediates Trypanosoma cruzi establishment by controlling the microbiota. Biochimie., vol. 123, p. 138-143, 2016.
ISSN: 0300-9084
Copyright: restricted access
Appears in Collections:MG - IRR - Artigos de Periódicos

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