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Imaging of the DNA damage-induced dynamics of nuclear proteins via nonlinear photoperturbation

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2013

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http://nbn-resolving.de/urn:nbn:de:bsz:352-216952
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https://doi.org/10.1002/jbio.201200170
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Journal of Biophotonics. 2013, 6(8), pp. 645-655. ISSN 1864-063X. eISSN 1864-0648. Available under: doi: 10.1002/jbio.201200170

Zusammenfassung

Understanding the cellular response to DNA strand breaks is crucial to decipher the mechanisms maintaining the integrity of our genome. We present a novel method to visualize how the mobility of nuclear proteins changes in response to localized DNA damage. DNA strand breaks are induced via nonlinear excitation with femtosecond laser pulses at λ = 1050 nm in a 3D-confined subnuclear volume. After a time delay of choice, protein mobility within this volume is analysed by two-photon photoactivation of PA-GFP fusion proteins at λ = 775 nm. By changing the position of the photoactivation spot with respect to the zone of lesion the influence of chromatin structure and of the distance from damage are investigated. As first applications we demonstrate a locally confined, time-dependent mobility increase of histone H1.2, and a progressive retardation of the DNA repair factor XRCC1 at damaged sites. This assay can be used to map the response of nuclear proteins to DNA damage in time and space.

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ISO 690TOMAS, Martin, Philipp BLUMHARDT, Anja DEUTZMANN, Tobias SCHWARZ, Dimitri KROMM, Alfred LEITENSTORFER, Elisa FERRANDO-MAY, 2013. Imaging of the DNA damage-induced dynamics of nuclear proteins via nonlinear photoperturbation. In: Journal of Biophotonics. 2013, 6(8), pp. 645-655. ISSN 1864-063X. eISSN 1864-0648. Available under: doi: 10.1002/jbio.201200170
BibTex
@article{Tomas2013-08Imagi-21695,
  year={2013},
  doi={10.1002/jbio.201200170},
  title={Imaging of the DNA damage-induced dynamics of nuclear proteins via nonlinear photoperturbation},
  number={8},
  volume={6},
  issn={1864-063X},
  journal={Journal of Biophotonics},
  pages={645--655},
  author={Tomas, Martin and Blumhardt, Philipp and Deutzmann, Anja and Schwarz, Tobias and Kromm, Dimitri and Leitenstorfer, Alfred and Ferrando-May, Elisa}
}
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