Caracterização funcional de VPS25, da via ESCRT, em Aspergillus fumigatus

Abstract

Aspergillus fumigatus is a filamentous, ubiquitous, and saprophytic fungus that causes invasive pulmonary aspergillosis, a disease of major medical importance, as it affects immunocompromised individuals, often leading to death. Intercellular communication is crucial for both microorganism physiology and pathogen-host interactions. In this context, intercellular communication mediated by the Endosomal Sorting Complex Required for Transport (ESCRT) pathway, conserved across eukaryotes, plays a significant role. Therefore, the aim of the present work is the in silico and functional characterization of the ESCRT pathway in Aspergillus fumigatus, through the generation of knockout strains for key ESCRT genes. To achieve this, all proteins orthologous to those in the ESCRT pathways of S. cerevisiae and C. albicans were identified, except for the MVB12 protein. These proteins show conserved domains, suggesting they may perform the same functions. Additionally, the ESCRT pathway is active during the vegetative development of A. fumigatus under different nutritional conditions (minimal and complete media). Furthermore, we generated a knockout strain for the vps25 gene. Functional characterization of this knockout strain demonstrated that VPS25 modulates both vegetative and reproductive pathways of the fungus and is involved in the biosynthesis, remodeling and integrity of A. fumigatus cell wall. Deletion of vps25 also affected factors of virulence, such as biofilm production, stress response, and recognition of A. fumigatus by phagocytes, leading to enhanced fungal clearance in vitro. Altogether, these results show that the ESCRT pathway, represented by VPS25, is active and plays a physiological role in A. fumigatus.