Spatio-temporal analysis of DNA damage repair using the X-ray microbeam |
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Authors: | G Schettino M Ghita and K M Prise |
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Institution: | (1) School of Nuclear Engineering, Purdue University, 400 Central Drive, West Lafayette, IN 47907, USA; |
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Abstract: | Cellular response to radiation damage is made by a complex network of
pathways and feedback loops whose spatiotemporal organization is still
unclear despite its decisive role in determining the fate of the damaged
cell. The single-cell approach and the high spatial resolution offered by
microbeams provide the perfect tool to study and quantify the dynamic
processes associated with the induction and repair of DNA damage. The soft
X-ray microbeam has been used to follow the development of radiation induced
foci in live cells by monitoring their size and intensity as a function of
dose and time using yellow fluorescent protein (YFP) tagging techniques.
Preliminary data indicate a delayed and linear rising of the intensity
signal indicating a slow kinetic for the accumulation of DNA repair protein
53BP1. A slow and limited foci diffusion has also been observed. Further
investigations are required to assess whatever such diffusion is consistent
with a random walk pattern or if it is the result of a more structured
lesion processing phenomenon. In conclusion, our data indicates that the use
of microbeams coupled to live cell microscopy represent a sophisticated
approach for visualizing and quantifying the dynamics changes of DNA
proteins at the damaged sites. |
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Keywords: | |
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