Autor(es): Marcos, Caroline Maria [UNESP] ; de Oliveira, Haroldo Cesar [UNESP] ; Assato, Patrícia Akemi [UNESP] ; Castelli, Rafael Fernando ; Fusco-Almeida, Ana Marisa [UNESP] ; Mendes-Giannini, Maria José Soares [UNESP]
Data: 2022
Identificador Persistente: http://hdl.handle.net/11449/233730
Origem: Oasisbr
Assunto(s): Antisense-RNA silencing; Cell wall; Dimorphism regulating-histidine kinase; Drk1; Paracoccidioides brasiliensis; Virulence
Made available in DSpace on 2022-05-01T09:47:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-10-01
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
P. brasiliensis is a thermally dimorphic fungus belonging to Paracoccidioides complex, causative of a systemic, endemic mycosis limited to Latin American countries. Signal transduc-tion pathways related to important aspects as surviving, proliferation according to the biological niches are linked to the fungal pathogenicity in many species, but its elucidation in P. brasiliensis remains poorly explored. As Drk1, a hybrid histidine kinase, plays regulators functions in other dimorphic fungi species, mainly in dimorphism and virulence, here we investigated its importance in P. brasilensis. We, therefore generated the respective recombinant protein, anti-PbDrk1 polyclonal antibody and a silenced strain. The Drk1 protein shows a random distribution including cell wall location that change its pattern during osmotic stress condition; moreover the P. brasiliensis treatment with anti-PbDrk1 antibody, which does not modify the fungus’s viability, resulted in decreased virulence in G. mellonella model and reduced interaction with pneumocytes. Down-regulating Pb-DRK1 yielded phenotypic alterations such as yeast cells with more elongated morphology, virulence attenuation in G. mellonella infection model, lower amount of chitin content, increased resistance to osmotic and cell wall stresses, and also caspofungin, and finally increased sensitivity to itraconazole. These observations highlight the importance of PbDrk1 to P. brasiliensis virulence, stress adaptation, morphology, and cell wall organization, and therefore it an interesting target that could help develop new antifungals.
School of Pharmaceutical Sciences São Paulo State University (UNESP)
Instituto Carlos Chagas Fundação Oswaldo Cruz (Fiocruz)
Laboratório Central de Multiusuários Faculdade de Ciências Agronômicas Campus Botucatu UNESP—Universidade Estadual Paulista
Programa de Pós-Graduação em Biologia Parasitária Instituto Oswaldo Cruz Fundação Oswaldo Cruz (Fiocruz)
School of Pharmaceutical Sciences São Paulo State University (UNESP)
Laboratório Central de Multiusuários Faculdade de Ciências Agronômicas Campus Botucatu UNESP—Universidade Estadual Paulista
CNPq: 164217/2020-7
FAPESP: 2015/03700-9
FAPESP: 2015/14023-8
FAPESP: 2016/17048-4
CNPq: 403586/2012-7
CAPES: 88881.141306/2017-01
