Mann Noah, Hill Jahiem, Wang Kenneth, Hughes Robert M
Department of Chemistry, East Carolina University, Greenville, North Carolina, United States.
Department of Chemistry, Davidson College, Davidson, North Carolina, United States.
bioRxiv. 2023 Dec 19:2023.10.04.560945. doi: 10.1101/2023.10.04.560945.
The actin cytoskeleton is a biosensor of cellular stress and a potential prognosticator of human disease. In particular, aberrant cytoskeletal structures such as cofilin-actin rods and stress granules formed in response to energetic and oxidative stress are closely linked to neurodegenerative diseases such as Alzheimer's, Parkinson's, and ALS. Whether these cytoskeletal phenomena can be harnessed for the development of biosensors for cytoskeletal dysfunction and, by extension, neurodegenerative disease progression, remains an open question. In this work, we describe the design and development of an optogenetic iteration of profilin, an actin monomer binding protein with critical functions in cytoskeletal dynamics. We demonstrate that this optically activated profilin ('OptoProfilin') can act as an optically triggered biosensor of applied cellular stress in select immortalized cell lines. Notably, OptoProfilin is a single component biosensor, likely increasing its utility for experimentalists. While a large body of preexisting work closely links profilin activity with cellular stress and neurodegenerative disease, this, to our knowledge, is the first example of profilin as an optogenetic biosensor of stress-induced changes in the cytoskeleton.
肌动蛋白细胞骨架是细胞应激的生物传感器,也是人类疾病的潜在预后指标。特别是,诸如因能量和氧化应激而形成的丝切蛋白 - 肌动蛋白杆和应激颗粒等异常细胞骨架结构与阿尔茨海默病、帕金森病和肌萎缩侧索硬化症等神经退行性疾病密切相关。这些细胞骨架现象是否可用于开发针对细胞骨架功能障碍以及由此延伸的神经退行性疾病进展的生物传感器,仍然是一个悬而未决的问题。在这项工作中,我们描述了一种肌动蛋白单体结合蛋白(在细胞骨架动力学中具有关键功能)—— 肌动蛋白结合蛋白(profilin)的光遗传学迭代的设计与开发。我们证明,这种光激活的肌动蛋白结合蛋白(“光遗传学肌动蛋白结合蛋白”)可以在选定的永生化细胞系中作为施加细胞应激的光触发生物传感器。值得注意的是,光遗传学肌动蛋白结合蛋白是一种单组分生物传感器,这可能会增加其对实验人员的实用性。虽然大量已有的研究将肌动蛋白结合蛋白的活性与细胞应激和神经退行性疾病紧密联系起来,但据我们所知,这是肌动蛋白结合蛋白作为应激诱导的细胞骨架变化的光遗传学生物传感器的首个实例。