Advanced search
Start date
Betweenand
Related content
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

A Kazal-type inhibitor is modulated by Trypanosoma cruzi to control microbiota inside the anterior midgut of Rhodnius prolixus

Full text
Author(s):
Soares, Tatiane S. [1] ; Buarque, Diego S. [1] ; Queiroz, Bruna R. [1] ; Gomes, Cicera M. [1] ; Braz, Gloria R. C. [2] ; Araujo, Ricardo N. [3] ; Pereira, Marcos H. [3] ; Guarneri, Alessandra A. [4] ; Tanaka, Aparecida S. [1]
Total Authors: 9
Affiliation:
[1] Univ Fed Sao Paulo, Dept Biochem, Escola Paulista Med, BR-04044020 Sao Paulo, SP - Brazil
[2] Univ Fed Rio de Janeiro, Inst Quim, Dept Biochem, BR-21941909 Rio De Janeiro, RJ - Brazil
[3] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Parasitol, BR-31270901 Belo Horizonte, MG - Brazil
[4] Fiocruz MS, Ctr Pesquisas Rene Rachou, BR-30190002 Belo Horizonte, MG - Brazil
Total Affiliations: 4
Document type: Journal article
Source: Biochimie; v. 112, p. 41-48, MAY 2015.
Web of Science Citations: 13
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 Rhodnius prolixus, named RpTI, which may be involved in microbiota - Trypanosoma 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 Trypanosoma 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 (K-i = 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 Trypanosoma cruzi trypomastigotes. The RpTI transcript was also knocked down by RNA interference prior to infection of Rhodnius prolixus with Trypanosoma cruzi. The amount of Trypanosoma 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 Rhodnius prolixus compared to non-silenced and infected insects. Our results suggest that Trypanosoma cruzi infection increases the expression of RpTI to mediate microbiota modulation and is important for parasite immediately after infection with Rhodnius prolixus. (C) 2015 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved. (AU)

FAPESP's process: 12/03657-8 - Inhibitor and proteases of ectoparasites: relationship of structure-function and identification of the role of these molecules in the interaction of diseases vector e their etiological agents
Grantee:Aparecida Sadae Tanaka
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 12/00723-0 - Interactions of three molecules from Triatoma infestans midgut with the protozoan Trypanosoma cruzi: an insight of Chagas disease vector- parasite relationships
Grantee:Diego de Souza Buarque
Support Opportunities: Scholarships in Brazil - Post-Doctoral