Caracterização molecular da proteína histidina quinase de Paracoccidioides brasiliensis

Detalhes bibliográficos
Ano de defesa: 2019
Autor(a) principal: Santos, Dener Lucas Araújo dos lattes
Orientador(a): Paccez, Juliano Domiraci lattes
Banca de defesa: Paccez, Juliano Domiraci, Bailão, Alexandre Melo, Tomazett, Mariana Vieira
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Goiás
Programa de Pós-Graduação: Programa de Pós-graduação em Genética e Biologia Molecular (ICB)
Departamento: Instituto de Ciências Biológicas - ICB (RG)
País: Brasil
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://repositorio.bc.ufg.br/tede/handle/tede/9457
Resumo: Prokaryotic and eukaryotic organisms (except those belonging to the phylum metazoa) have developed a system named phosphorelay, which is responsible for adaptation to different environmental conditions such as oxidative stress, osmotic stress and thermal shock. Studies have demonstrated that inhibition of hybrid histidine kinase, the stimulus sensory protein in the phosphorelay pathway, affects morphological transition from mycelial fungi to yeast. Among the thermodymorphic fungi, Paracoccidioides spp. are among the most prevalent in Latin America, mainly in Brazil, as the cause of the systemic mycosis paracoccidioidomycosis (PCM). The exposure of fungi from this complex to the host body temperature (37 ° C), leads to the transition from the mycelial to the parasitic yeast form. It is notable that the morphological transition is necessary for the pathogenicity of Paracoccidioides spp.. Thus, the objective of this study is to characterize the hybrid histidine kinase (HHK) protein and to evaluate its level of expression during the dimorphism of Paracoccidioides brasiliensis. We carried out similarity analyzes that indicated that HHK is highly conserved in other dimorphic fungi such as H. capsulatum and B. dermatitidis. In addition, a region of the HHK gene was cloned into pET32a expression vector and the recombinant protein was generated. After confirming the identity of HHK in UPLC-MSE, we produced polyclonal anti-histidine kinase antibodies. RT-qPCR revealed that PbHHK is expressed in greater amounts transition when compared to the other morphological phases. In conclusion, the high conservation of histidine kinase in dimorphic fungi combined with their increased expression in the transition phase demonstrates the importance of PbHHK for the morphological transition of P. brasiliensis.