Instituto Carlos Chagas, Fundação Oswaldo Cruz (Fiocruz), Curitiba, Brazil.
Programa de Pós-Graduação em Biologia Parasitária, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
Infect Immun. 2024 Apr 9;92(4):e0003724. doi: 10.1128/iai.00037-24. Epub 2024 Mar 12.
Small molecules are components of fungal extracellular vesicles (EVs), but their biological roles are only superficially known. is a eukaryotic gene that is required for the activity of benzimidazoles against . In this study, during the phenotypic characterization of mutants expected to lack expression, we observed a reduced EV production. Whole-genome sequencing, RNA-Seq, and cellular proteomics revealed that, contrary to our initial findings, these mutants expressed Nop16 but exhibited altered expression of 14 genes potentially involved in sugar transport. Based on this observation, we designated these mutant strains as Past1 and Past2, representing otentially ltered ugar ransport. Analysis of the small molecule composition of EVs produced by wild-type cells and the Past1 and Past2 mutant strains revealed not only a reduced number of EVs but also an altered small molecule composition. In a model of infection, the Past1 and Past2 mutant strains were hypovirulent. The hypovirulent phenotype was reverted when EVs produced by wild-type cells, but not mutant EVs, were co-injected with the mutant cells in . These results connect EV biogenesis, cargo, and cryptococcal virulence.
小分子是真菌细胞外囊泡(EVs)的组成部分,但它们的生物学功能知之甚少。 is a eukaryotic gene 是一种真核基因,对 针对 的苯并咪唑类药物的活性是必需的。在这项研究中,在对预期缺乏 表达的 突变体进行表型特征分析时,我们观察到 EV 产生减少。全基因组测序、RNA-Seq 和细胞蛋白质组学揭示,与我们最初的发现相反,这些突变体表达了 Nop16,但表现出 14 个可能参与糖转运的基因表达改变。基于这一观察结果,我们将这些突变菌株命名为 Past1 和 Past2,代表可能改变的糖转运。对野生型细胞和 Past1 和 Past2 突变菌株产生的 EV 中小分子组成的分析不仅揭示了 EV 数量减少,还揭示了小分子组成的改变。在感染模型中,Past1 和 Past2 突变株的毒力降低。当将野生型细胞产生的 EV 而不是突变型 EV 与 中的突变细胞共注射时,Past1 和 Past2 突变株的低毒力表型得到逆转。这些结果将 EV 生物发生、货物和隐球菌毒力联系起来。