Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, United States of America.
Institute of Molecular Infection Biology, University of Würzburg, Würzburg, Germany.
PLoS Pathog. 2023 Sep 28;19(9):e1011692. doi: 10.1371/journal.ppat.1011692. eCollection 2023 Sep.
The signals that denote mammalian host environments and dictate the activation of signaling pathways in human-associated microorganisms are often unknown. The transcription regulator Rtg1/3 in the human fungal pathogen Candida albicans is a crucial determinant of host colonization and pathogenicity. Rtg1/3's activity is controlled, in part, by shuttling the regulator between the cytoplasm and nucleus of the fungus. The host signal(s) that Rtg1/3 respond(s) to, however, have remained unclear. Here we report that neutrophil-derived reactive oxygen species (ROS) direct the subcellular localization of this C. albicans transcription regulator. Upon engulfment of Candida cells by human or mouse neutrophils, the regulator shuttles to the fungal nucleus. Using genetic and chemical approaches to disrupt the neutrophils' oxidative burst, we establish that the oxidants produced by the NOX2 complex-but not the oxidants generated by myeloperoxidase-trigger Rtg1/3's migration to the nucleus. Furthermore, screening a collection of C. albicans kinase deletion mutants, we implicate the MKC1 signaling pathway in the ROS-dependent regulation of Rtg1/3 in this fungus. Finally, we show that Rtg1/3 contributes to C. albicans virulence in the nematode Caenorhabditis elegans in an ROS-dependent manner as the rtg1 and rtg3 mutants display virulence defects in wild-type but not in ROS deficient worms. Our findings establish NOX2-derived ROS as a key signal that directs the activity of the pleiotropic fungal regulator Rtg1/3.
哺乳动物宿主环境的信号以及决定人类相关微生物信号通路激活的信号往往是未知的。人类真菌病原体白色念珠菌中的转录调节因子 Rtg1/3 是决定宿主定植和致病性的关键决定因素。Rtg1/3 的活性部分受其在真菌细胞质和细胞核之间穿梭的调控。然而,Rtg1/3 所响应的宿主信号仍不清楚。在这里,我们报告说中性粒细胞衍生的活性氧物质(ROS)指导这种白色念珠菌转录调节剂的亚细胞定位。当人类或鼠中性粒细胞吞噬白色念珠菌细胞时,该调节剂会穿梭到真菌核中。通过使用遗传和化学方法破坏中性粒细胞的氧化爆发,我们确定了由 NOX2 复合物产生的氧化剂-而不是由髓过氧化物酶产生的氧化剂-触发 Rtg1/3 向核内迁移。此外,通过筛选一组白色念珠菌激酶缺失突变体,我们发现 MKC1 信号通路参与了 ROS 依赖性调节 Rtg1/3 在该真菌中的作用。最后,我们表明 Rtg1/3 以 ROS 依赖性方式有助于白色念珠菌在秀丽隐杆线虫中的毒力,因为 rtg1 和 rtg3 突变体在野生型而非 ROS 缺陷型蠕虫中表现出毒力缺陷。我们的研究结果确立了 NOX2 衍生的 ROS 作为一种关键信号,指导多效性真菌调节因子 Rtg1/3 的活性。