The first characterization of a cystatin and a cathepsin L-like peptidase from Aedes aegypti and their possible role in DENV infection by the modulation of apoptosis

Autor(es)

Oliveira, Felipe A. A. ^[1] ; Buri, Marcus V. ^[1] ; Rodriguez, Boris L. ^[1] ; Costa-da-Silva, Andre L. ^[2] ; Araujo, Helena R. C. ^[2] ; Capurro, Margareth L. ^[2] ; Lu, Stephen ^[1] ; Tanaka, Aparecida S. ^[1]

Afiliação do(s) autor(es)

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

^[2] Univ Sao Paulo, Inst Ciencias Biomed, Dept Parasitol, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 146, p. 141-149, MAR 1 2020.
• Citações Web of Science: 0

Resumo

Recently, a salivary gland transcriptome study demonstrated that the transcripts of a putative cystatin gene (SeqID AAEL013287; Aacystatins) from Aedes aegypti were increased in DENV2-infected mosquitoes and that silencing of the Aacystatin gene resulted in an increase in DENV titres. In this work, Aacystatin was biochemically characterized; the purified recombinant inhibitor was able to inhibit typical cysteine proteases with a Ki in the nM range. Pulldown assays using Aag2 cell extracts identified a cathepsin L-like peptidase (AaCatL) as a possible target of Aacystatin. Purified recombinant AaCatL had an optimal pH of 5.0 and displayed a preference for Leu, Val and Phe residues at P2, which is common for other cathepsin L-like peptidases. Transcription analysis of Aacystatin and AaCatL in the salivary glands and midgut of DENV2-infected mosquitoes revealed a negative correlation between DENV2 titres and levels of the inhibitor and peptidase, suggesting their involvement in DENV2-mosquito interactions. Considering that apoptosis may play an important role during viral infections, the possible involvement of Aacystatin in staurosporine-induced apoptosis in Aag2 cells was investigated; the results showed higher expression of the inhibitor in treated cells; moreover, pre incubation with rAacystatin was able to increase Aag2 cell viability. (C) 2019 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 13/16609-4 - Conversão do sexo do Aedes aegypti por transgenia
• Beneficiário: Helena Corrêa de Araújo Gomes
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• 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