Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/12417
Title: A Kazal-type inhibitor is modulated by Trypanosoma cruzi to control microbiota inside the anterior midgut of Rhodnius prolixus.
Authors: Soares, Tatiane Sanches
Buarque, Diego de Souza
Queiroz, Bruna Regina
Gomes, Cícera Maria
Braz, Glória Regina Cardoso
Araújo, Ricardo Nascimento
Pereira, Marcos Horacio
Guarnieri, Alessandra Aparecida
Tanaka, Aparecida Sadae
Affilliation: Universidade Federal de São Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. São Paulo, SP, Brazil
Universidade Federal de São Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. São Paulo, SP, Brazil
Universidade Federal de São Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. São Paulo, SP, Brazil
Universidade Federal de São Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. São Paulo, SP, Brazil
Universidade Federal do Rio de Janeiro. Instituto de Química. Departamento de Bioquimica. Rio de Janeiro, RJ, Brazil
Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brazil
Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brazil
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brazil
Universidade Federal de São Paulo. Escola Paulista de Medicina. Departamento de Bioquimica. São Paulo, SP, Brazil
Abstract: The triatomine insect, Rhodnius prolixus, is a vector of Trypanosoma cruzi, a protozoan parasite that causes Chagas disease. The parasite must overcome immune response and microbiota to develop inside the midgut of triatomines. In this study, we expressed, purified and characterized a Kazal-type inhibitor from the midgut of R. prolixus, named RpTI, which may be involved in microbiota – T. cruzi interactions. The qPCR showed that the RpTI transcript was primarily expressed in tissues from the intestinal tract and that it was upregulated in the anterior midgut after T. cruzi infection. A 315-bp cDNA fragment encoding the mature protein was cloned into the pPIC9 vector and expressed in Pichia pastoris system. Recombinant RpTI (rRpTI) was purified on a trypsin-Sepharose column and had a molecular mass of 11.5 kDa as determined by SDS-PAGE analysis. This protein inhibited trypsin (Ki = 0.42 nM), whereas serine proteases from the coagulation cascade were not inhibited. Moreover, trypanocidal assays revealed that rRpTI did not interfere in the viability of T. cruzi trypomastigotes. The RpTI transcript was also knocked down by RNA interference prior to infection of R. prolixus with T. cruzi. The amount of T. cruzi in the anterior midgut was significantly lower in RpTI knockdown insects compared to the non-silenced groups. We also verified that the bacterial load is higher in the anterior midgut of silenced and infected R. prolixus compared to non-silenced and infected insects. Our results suggest that T. cruzi infection increases the expression of RpTI to mediate microbiota modulation and is important for parasite immediately after infection with R. prolixus
Keywords: Anterior midgut
Kazal family
Microbiota
Rhodnius prolixus
Trypanosoma cruzi
Issue Date: 2015
Publisher: Elsevier Masson SAS
Citation: SOARES, Tatiane Sanches et al. A Kazal-type inhibitor is modulated by Trypanosoma cruzi to control microbiota inside the anterior midgut of Rhodnius prolixus. Biochimie, Vol. 112, p. 41–48, 2015.
DOI: 10.1016/j.biochi.2015.02.014
ISSN: 0300-9084
Copyright: restricted access
Appears in Collections:MG - IRR - Artigos de Periódicos




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