Friedman Nir, Vardi Shuki, Ronen Michal, Alon Uri, Stavans Joel
Department of Physics of Complex Systems, The Weizmann Institute of Science, Rehovot, Israel.
PLoS Biol. 2005 Jul;3(7):e238. doi: 10.1371/journal.pbio.0030238. Epub 2005 Jun 21.
The SOS genetic network is responsible for the repair/bypass of DNA damage in bacterial cells. While the initial stages of the response have been well characterized, less is known about the dynamics of the response after induction and its shutoff. To address this, we followed the response of the SOS network in living individual Escherichia coli cells. The promoter activity (PA) of SOS genes was monitored using fluorescent protein-promoter fusions, with high temporal resolution, after ultraviolet irradiation activation. We find a temporal pattern of discrete activity peaks masked in studies of cell populations. The number of peaks increases, while their amplitude reaches saturation, as the damage level is increased. Peak timing is highly precise from cell to cell and is independent of the stage in the cell cycle at the time of damage. Evidence is presented for the involvement of the umuDC operon in maintaining the pattern of PA and its temporal precision, providing further evidence for the role UmuD cleavage plays in effecting a timed pause during the SOS response, as previously proposed. The modulations in PA we observe share many features in common with the oscillatory behavior recently observed in a mammalian DNA damage response. Our results, which reveal a hitherto unknown modulation of the SOS response, underscore the importance of carrying out dynamic measurements at the level of individual living cells in order to unravel how a natural genetic network operates at the systems level.
SOS 基因网络负责细菌细胞中 DNA 损伤的修复/旁路。虽然该反应的初始阶段已得到充分表征,但对于诱导后反应的动力学及其关闭过程了解较少。为了解决这个问题,我们追踪了单个活的大肠杆菌细胞中 SOS 网络的反应。在紫外线照射激活后,使用荧光蛋白-启动子融合物以高时间分辨率监测 SOS 基因的启动子活性(PA)。我们发现了在细胞群体研究中被掩盖的离散活性峰的时间模式。随着损伤水平的增加,峰的数量增加,而其幅度达到饱和。峰的时间在细胞之间高度精确,并且与损伤时细胞周期的阶段无关。有证据表明 umuDC 操纵子参与维持 PA 的模式及其时间精度,为先前提出的 UmuD 切割在 SOS 反应期间实现定时暂停中所起的作用提供了进一步的证据。我们观察到的 PA 调制与最近在哺乳动物 DNA 损伤反应中观察到的振荡行为有许多共同特征。我们的结果揭示了 SOS 反应迄今未知的调制,强调了在单个活细胞水平上进行动态测量以揭示自然遗传网络在系统水平上如何运作的重要性。