Department of Microbiology & Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
Department of Ophthalmology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America.
PLoS Pathog. 2023 Oct 31;19(10):e1011435. doi: 10.1371/journal.ppat.1011435. eCollection 2023 Oct.
The Aspergillus fumigatus unfolded protein response (UPR) is a two-component relay consisting of the ER-bound IreA protein, which splices and activates the mRNA of the transcription factor HacA. Spliced hacA accumulates under conditions of acute ER stress in vitro, and UPR null mutants are hypovirulent in a murine model of invasive pulmonary infection. In this report, we demonstrate that a hacA deletion mutant (ΔhacA) is furthermore avirulent in a model of fungal keratitis, a corneal infection, and an important cause of ocular morbidity and unilateral blindness worldwide. Interestingly, we demonstrate that A. fumigatus hacA is spliced in infected lung samples, but not in the cornea, suggesting the amount of ER stress experienced by the fungus varies upon the host niche. To better understand how the UPR contributes to fungal cell biology across a spectrum of ER-stress levels, we employed transcriptomics on the wild-type and ΔhacA strains in glucose minimal media (low stress), glucose minimal media with dithiothreitol (high stress), and gelatin minimal media as a proxy for the nutrient stress encountered in the cornea (mid-level stress). These data altogether reveal a unique HacA-dependent transcriptome under each condition, suggesting that HacA activity is finely-tuned and required for proper fungal adaptation in each environment. Taken together, our results indicate that the fungal UPR could serve as an important antifungal target in the setting of both invasive pulmonary and corneal infections.
烟曲霉 unfolded 蛋白反应 (UPR) 是一个由两个组件组成的接力系统,包括内质网结合的 IreA 蛋白,它剪接并激活转录因子 HacA 的 mRNA。在体外急性内质网应激条件下,剪接的 hacA 积累,而 UPR 缺失突变体在侵袭性肺部感染的小鼠模型中毒力降低。在本报告中,我们证明了 hacA 缺失突变体 (ΔhacA) 在真菌角膜炎模型中也具有毒力缺失,这是一种角膜感染,也是全球眼部发病率和单侧失明的重要原因。有趣的是,我们证明烟曲霉 hacA 在感染的肺部样本中被剪接,但在角膜中没有,这表明真菌经历的内质网应激量因宿主小生境而异。为了更好地了解 UPR 如何在一系列内质网应激水平下影响真菌细胞生物学,我们在葡萄糖最低培养基(低应激)、葡萄糖最低培养基加二硫苏糖醇(高应激)和明胶最低培养基(作为角膜中遇到的营养应激的替代物)中对野生型和 ΔhacA 菌株进行了转录组学研究。这些数据共同揭示了每种条件下独特的依赖 HacA 的转录组,表明 HacA 活性被精细调节,并且在每种环境中都需要适当的真菌适应。总之,我们的结果表明,真菌 UPR 可以作为侵袭性肺部和角膜感染的重要抗真菌靶点。