A new antimicrobial protein from the anterior midgut of Triatoma infestans mediates Trypanosoma cruzi establishment by controlling the microbiota

Autor(es)

Buarque, Diego S. ^[1] ; Gomes, Cicera M. ^[1] ; Araujo, Ricardo N. ^[2] ; Pereira, Marcos H. ^[2] ; Ferreira, Roberta C. ^[3] ; Guarneri, Alessandra A. ^[3] ; Tanaka, Aparecida S. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Biochem, BR-04044020 Sao Paulo, SP - Brazil

^[2] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Parasitol, BR-31270901 Belo Horizonte, MG - Brazil

^[3] Fiocruz MS, Ctr Pesquisas Rene Rachou, BR-30190002 Belo Horizonte, MG - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biochimie; v. 123, p. 138-143, APR 2016.
• Citações Web of Science: 6

Resumo

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. (C) 2016 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved. (AU)

• Processo FAPESP: 12/03657-8 - Inibidores e proteases de ectoparasitas: relação de estrutura-função e identificação do papel dessas moléculas na interação de vetores de doenças e seus agentes etiológicos
• Beneficiário: Aparecida Sadae Tanaka
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 12/00723-0 - Estudo da interação de três moléculas do trato digestivo do inseto Triatoma infestans com o protozoário Trypanosoma cruzi: uma visão da relação parasita - vetor da Doença de Chagas
• Beneficiário: Diego de Souza Buarque
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado