Murciano-Julià Guillem, Francos-Cárdenas Marina, Salat-Canela Clàudia, Hidalgo Elena, Ayté José
Oxidative Stress and Cell Cycle Group, Universitat Pompeu Fabra, Barcelona, Spain.
PLoS Biol. 2025 Jan 7;23(1):e3002969. doi: 10.1371/journal.pbio.3002969. eCollection 2025 Jan.
Fission yeast is an excellent model system that has been widely used to study the mechanism that control cell cycle progression. However, there is a lack of tools that allow to measure with high precision the duration of the different phases of the cell cycle in individual cells. To circumvent this problem, we have developed a fluorescent reporter that allows the quantification of the different phases of the cell cycle at the single-cell level in most genetic backgrounds. To prove the accuracy of this fluorescent reporter, we have tested the reporter in strains known to have a delay in the G1/S or G2/M transitions, confirming the strength and versatility of the system. An advantage of this reporter is that it eliminates the need for culture synchronization, avoiding stressing the cells. Using this reporter, we show that unperturbed cells lacking Sty1 have a standard cell cycle length and distribution and that the extended length of these cells is due to their increased cell growth rate but not to alterations in their cell cycle progression.
裂殖酵母是一种优秀的模型系统，已被广泛用于研究控制细胞周期进程的机制。然而，缺乏能够高精度测量单个细胞中细胞周期不同阶段持续时间的工具。为了解决这个问题，我们开发了一种荧光报告基因，它能够在大多数遗传背景下在单细胞水平上对细胞周期的不同阶段进行定量分析。为了证明这种荧光报告基因的准确性，我们在已知在G1/S或G2/M转换中存在延迟的菌株中测试了该报告基因，证实了该系统的优势和通用性。这种报告基因的一个优点是它无需进行培养同步化，避免了对细胞造成压力。使用这个报告基因，我们表明缺乏Sty1的未受干扰的细胞具有标准的细胞周期长度和分布，并且这些细胞延长的长度是由于它们增加的细胞生长速率，而不是由于它们细胞周期进程的改变。
