Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210.
The Ohio State Biochemistry Program, The Ohio State University, Columbus, OH 43210.
Mol Biol Cell. 2021 Jun 1;32(12):1181-1192. doi: 10.1091/mbc.E20-11-0734. Epub 2021 Apr 7.
After growing on surfaces, including those of medical and industrial importance, fungal biofilms self-generate internal microenvironments. We previously reported that gaseous microenvironments around founder cells change during biofilm formation causing microtubules to disassemble under control of the hypoxic transcription factor SrbA. Here we investigate if biofilm formation might also promote changes to structures involved in exocytosis and endocytosis. During biofilm formation, the endoplasmic reticulum (ER) remained intact but ER exit sites and the Golgi apparatus were modified as were endocytic actin patches. The biofilm-driven changes required the SrbA hypoxic transcription factor and could be triggered by nitric oxide, further implicating gaseous regulation of biofilm cellular architecture. By tracking green fluorescent protein (GFP)-Atg8 dynamics, biofilm founder cells were also observed to undergo autophagy. Most notably, biofilm cells that had undergone autophagy were triggered into further autophagy by spinning disk confocal light. Our findings indicate that fungal biofilm formation modifies the secretory and endocytic apparatus and show that biofilm cells can also undergo autophagy that is reactivated by light. The findings provide new insights into the changes occurring in fungal biofilm cell biology that potentially impact their unique characteristics, including antifungal drug resistance.
在包括医学和工业重要性的表面上生长后,真菌生物膜会自我生成内部微环境。我们之前曾报道过,创始细胞周围的气态微环境在生物膜形成过程中会发生变化,导致微管在缺氧转录因子 SrbA 的控制下解体。在这里,我们研究了生物膜形成是否也可能促进参与胞吐作用和胞吞作用的结构发生变化。在生物膜形成过程中,内质网 (ER) 保持完整,但 ER 出口位点和高尔基体被修饰,内吞作用的肌动蛋白斑也被修饰。生物膜驱动的变化需要 SrbA 缺氧转录因子,并且可以被一氧化氮触发,这进一步表明气态调节生物膜细胞结构。通过跟踪绿色荧光蛋白 (GFP)-Atg8 的动态变化,还观察到生物膜创始细胞经历自噬。值得注意的是,通过旋转盘共聚焦光,经历自噬的生物膜细胞被进一步触发自噬。我们的研究结果表明,真菌生物膜形成会改变分泌和内吞装置,并表明生物膜细胞也可以经历自噬,这种自噬可以被光重新激活。这些发现为真菌生物膜细胞生物学中发生的变化提供了新的见解,这些变化可能会影响它们的独特特征,包括抗真菌药物耐药性。