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The p53 response in single cells is linearly correlated to the number of DNA breaks without a distinct threshold

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Item Type:Article
Title:The p53 response in single cells is linearly correlated to the number of DNA breaks without a distinct threshold
Creators Name:Loewer, A. and Karanam, K. and Mock, C. and Lahav, G.
Abstract:BACKGROUND: The tumor suppressor protein p53 is activated by cellular stress. DNA double strand breaks (DSBs) induce the activation of the kinase ATM, which stabilizes p53 and activates its transcriptional activity. Single cell analysis revealed that DSBs induced by gamma irradiation triggers p53 accumulation in a series of pulses that vary in number from cell to cell. Higher levels of irradiation increases the number of p53 pulses suggesting that they arise from periodic examination of the damage by ATM. If damage persists, additional pulses of p53 are triggered. The threshold of damage required for activating a p53 pulse is unclear. Previous studies that averaged the response across cell populations suggested that one or two DNA breaks are sufficient for activating ATM and p53. However, it is possible that by averaging over a population of cells important features of the dependency between DNA breaks and p53 dynamics are missed. RESULTS: Using fluorescent reporters we developed a system for following in individual cells the number of DSBs, the kinetics of repair and the p53 response. We found a large variation in the initial number of DSBs and the rate of repair between individual cells. Cells with higher number of DSBs had higher probability of showing a p53 pulse. However, there was no distinct threshold number of breaks for inducing a p53 pulse. We present evidence that the decision to activate p53 given a specific number of breaks is not entirely stochastic, but instead is influenced by both cell-intrinsic factors and previous exposure to DNA damage. We also show that the natural variations in the initial amount of p53, rate of DSB repair and cell cycle phase do not affect the probability of activating p53 in response to DNA damage. CONCLUSIONS: The use of fluorescent reporters to quantify DNA damage and p53 levels in live cells provided a quantitative analysis of the complex interrelationships between both processes. Our study shows that p53 activation differs even between cells that have a similar number of DNA breaks. Understanding the origin and consequences of such variability in normal and cancerous cells is crucial for developing efficient and selective therapeutic interventions.
Keywords:Double Strand Breaks, p53, Live Imaging, Single Cells, Pulses
Source:BMC Biology
Publisher:BioMed Central
Page Range:114
Date:19 November 2013
Official Publication:https://doi.org/10.1186/1741-7007-11-114
PubMed:View item in PubMed

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