Zhang Jiantao, Benko Zsigmond, Zhang Chenyu, Zhao Richard Y
Department of Pathology, School of Medicine, University of Maryland, Baltimore, MD 21201, USA.
Department of Molecular Biotechnology and Microbiology, Faculty of Science and Technology, University of Debrecen, 4032 Debrecen, Hungary.
Pathogens. 2024 Jul 4;13(7):566. doi: 10.3390/pathogens13070566.
Fission yeast, a single-cell eukaryotic organism, shares many fundamental cellular processes with higher eukaryotes, including gene transcription and regulation, cell cycle regulation, vesicular transport and membrane trafficking, and cell death resulting from the cellular stress response. As a result, fission yeast has proven to be a versatile model organism for studying human physiology and diseases such as cell cycle dysregulation and cancer, as well as autophagy and neurodegenerative diseases like Alzheimer's, Parkinson's, and Huntington's diseases. Given that viruses are obligate intracellular parasites that rely on host cellular machinery to replicate and produce, fission yeast could serve as a surrogate to identify viral proteins that affect host cellular processes. This approach could facilitate the study of virus-host interactions and help identify potential viral targets for antiviral therapy. Using fission yeast for functional characterization of viral genomes offers several advantages, including a well-characterized and haploid genome, robustness, cost-effectiveness, ease of maintenance, and rapid doubling time. Therefore, fission yeast emerges as a valuable surrogate system for rapid and comprehensive functional characterization of viral proteins, aiding in the identification of therapeutic antiviral targets or viral proteins that impact highly conserved host cellular functions with significant virologic implications. Importantly, this approach has a proven track record of success in studying various human and plant viruses. In this protocol, we present a streamlined and scalable molecular cloning strategy tailored for genome-wide and comprehensive functional characterization of viral proteins in fission yeast.
裂殖酵母是一种单细胞真核生物，与高等真核生物共享许多基本的细胞过程，包括基因转录与调控、细胞周期调控、囊泡运输和膜 trafficking，以及细胞应激反应导致的细胞死亡。因此，裂殖酵母已被证明是一种通用的模式生物，可用于研究人类生理学和疾病，如细胞周期失调和癌症，以及自噬和神经退行性疾病，如阿尔茨海默病、帕金森病和亨廷顿病。鉴于病毒是专性细胞内寄生虫，依赖宿主细胞机制进行复制和产生，裂殖酵母可作为一种替代物，用于鉴定影响宿主细胞过程的病毒蛋白。这种方法有助于研究病毒-宿主相互作用，并有助于确定抗病毒治疗的潜在病毒靶点。使用裂殖酵母对病毒基因组进行功能表征具有几个优点，包括特征明确的单倍体基因组稳健性、成本效益、易于维护和快速倍增时间。因此，裂殖酵母成为一种有价值的替代系统，可用于快速、全面地对病毒蛋白进行功能表征，有助于鉴定治疗性抗病毒靶点或影响具有重大病毒学意义的高度保守宿主细胞功能的病毒蛋白。重要的是，这种方法在研究各种人类和植物病毒方面有成功的记录。在本方案中，我们提出了一种简化且可扩展的分子克隆策略，专门用于在裂殖酵母中对病毒蛋白进行全基因组和全面的功能表征。
