Silva Lana O'Hara Souza, Moreira Thalison Rodrigues, Gonçales Relber Aguiar, Tomazett Mariana Vieira, Parente-Rocha Juliana Alves, Mattos Karine, Paccez Juliano Domiraci, Ruiz Orville Hernandez, Pereira Maristela, Soares Célia Maria de Almeida, Weber Simone Schneider, Cruz-Leite Vanessa Rafaela Milhomem, Borges Clayton Luiz
Laboratory of Molecular Biology, Institute of Biological Sciences, Federal University of Goiás, Goiânia 74690-900, Brazil.
Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4700-000 Braga, Portugal.
Microorganisms. 2022 Oct 12;10(10):2011. doi: 10.3390/microorganisms10102011.
Nitrogen is a crucial nutrient for microorganisms that compose essential biomolecules. However, hosts limit this nutrient as a strategy to counter infections, therefore, pathogens use adaptive mechanisms to uptake nitrogen from alternative sources. In fungi, nitrogen catabolite repression (NCR) activates transcription factors to acquire nitrogen from alternative sources when preferential sources are absent. Formamidase has been related to nitrogen depletion in through formamide degradation to use the released ammonia as a nitrogen source. In spp., formamidase is highly expressed in transcriptomic and proteomic analyses. Here, we aim to investigate the importance of formamidase to . Thereby, we developed a silenced strain of gene (AsFmd) by antisense RNA technology using -mediated transformation (ATMT). The AsFmd strain led to increased urease expression, an enzyme related to nitrogen assimilation in other fungi, suggesting that might explore urease as an alternative route for ammonia metabolism as a nitrogen source. Moreover, formamidase was important for fungal survival inside macrophages, as fungal recovery after macrophage infection was lower in AsFmd compared to wild-type (WT) strain. Our findings suggest potential alternatives of nitrogen acquisition regulation in , evidencing formamidase influence in fungal virulence.
氮是构成重要生物分子的微生物的关键营养素。然而，宿主会限制这种营养素作为对抗感染的一种策略，因此，病原体利用适应性机制从替代来源摄取氮。在真菌中，氮分解代谢物阻遏（NCR）会激活转录因子，以便在缺乏优先来源时从替代来源获取氮。甲酰胺酶通过甲酰胺降解将释放的氨用作氮源，从而与氮消耗有关。在 种中，甲酰胺酶在转录组和蛋白质组分析中高度表达。在这里，我们旨在研究甲酰胺酶对 的重要性。因此，我们使用 介导的转化（ATMT）通过反义RNA技术开发了一种 基因（AsFmd）沉默菌株。AsFmd菌株导致脲酶表达增加，脲酶是一种与其他真菌中的氮同化相关的酶，这表明 可能将脲酶作为氨代谢作为氮源的替代途径。此外，甲酰胺酶对于巨噬细胞内的真菌存活很重要，因为与野生型（WT）菌株相比，AsFmd中巨噬细胞感染后的真菌回收率较低。我们的研究结果表明 在氮获取调节方面可能存在替代途径，证明了甲酰胺酶对真菌毒力的影响。
