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53BP1 and MDC1 foci formation in HT-1080 cells for low- and high-LET microbeam irradiations

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Abstract

An improved assessment of the biological effects and related risks of low doses of ionizing radiation is currently an important issue in radiation biology. Irradiations using microbeams are particularly well suited for precise and localized dose depositions, whereas recombinant cell lines with fluorescent proteins allow the live observation of radiation-induced foci. Living cells of the fibrosarcoma cell line HT-1080 stably expressing 53BP1 or full-length reconstituted MDC1 fused to Green Fluorescent Protein (GFP) were irradiated with protons and α-particles of linear energy transfers (LETs) of 15 and 75 keV/μm, respectively. Using a microbeam, the irradiations were carried out in line patterns, which facilitated the discrimination between undefined background and radiation-induced foci. As expected, foci formation and respective kinetics from α-particle irradiations with a high LET of 75 keV/μm could be detected in a reliable manner by both fusion proteins, as reported previously. Colocalization of γ-H2AX foci confirmed the DSB nature of the detected foci. As a novel result, the application of protons with low LET of 15 keV/μm generated 53BP1- and MDC1-mediated foci of almost equal size and slightly different kinetics. This new data expands the capability of 53BP1 and wild-type MDC1 on visible foci formation in living cells after irradiation with low-LET particles. Furthermore, the kinetics in HT-1080 cells for α-particle irradiation show a delay of about 20 s for 53BP1 foci detection compared to wild-type MDC1, confirming the hierarchical assembly of both proteins. Preliminary data for proton irradiations are shown and also these indicate a delay for 53BP1 versus MDC1.

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Acknowledgments

The authors would like to thank O. Döhr, H. Eggestein, T. Heldt, and M. Hoffmann for the operation of the PTB ion accelerators, K. Beverung, A. Heiske and S. Löb for their constant support with the irradiation of cells and S. Fähnrich and Y. Merkhoffer for cell culture and preparation of cells.

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Authors and Affiliations

  1. Physikalisch-Technische Bundesanstalt (PTB), Bundesallee 100, 38116, Braunschweig, Germany

    Marita Mosconi, Ulrich Giesen & Frank Langner

  2. Heinrich Heine University (HHU), 40225, Düsseldorf, Germany

    Christian Mielke & Ilaria Dalla Rosa

  3. Leibniz Institut Deutsche Sammlung für Mikroorganismen und Zellkulturen (DSMZ), 38124, Braunschweig, Germany

    Wilhelm G. Dirks

Authors
  1. Marita Mosconi
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  2. Ulrich Giesen
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  3. Frank Langner
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  4. Christian Mielke
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  5. Ilaria Dalla Rosa
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  6. Wilhelm G. Dirks
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Corresponding authors

Correspondence to Marita Mosconi or Ulrich Giesen.

Additional information

This paper is based on a presentation made at the 9th International Microbeam Workshop, July 15–17, 2010, in Darmstadt, Germany.

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Mosconi, M., Giesen, U., Langner, F. et al. 53BP1 and MDC1 foci formation in HT-1080 cells for low- and high-LET microbeam irradiations. Radiat Environ Biophys 50, 345–352 (2011). https://doi.org/10.1007/s00411-011-0366-9

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  • DOI: https://doi.org/10.1007/s00411-011-0366-9

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