TY - JOUR A1 - Eryilmaz, Marion A1 - Schmitt, Eberhard A1 - Krufczik, Matthias A1 - Theda, Franziska A1 - Lee, Jin-Ho A1 - Cremer, Christoph A1 - Bestvater, Felix A1 - Schaufler, Wladimir A1 - Hausmann, Michael A1 - Hildenbrand, Georg T1 - Localization Microscopy Analyses of MRE11 Clusters in 3D-Conserved Cell Nuclei of Different Cell Lines JF - Cancers N2 - In radiation biophysics, it is a subject of nowadays research to investigate DNA strand break repair in detail after damage induction by ionizing radiation. It is a subject of debate as to what makes up the cell’s decision to use a certain repair pathway and how the repair machinery recruited in repair foci is spatially and temporarily organized. Single-molecule localization microscopy (SMLM) allows super-resolution analysis by precise localization of single fluorescent molecule tags, resulting in nuclear structure analysis with a spatial resolution in the 10 nm regime. Here, we used SMLM to study MRE11 foci. MRE11 is one of three proteins involved in the MRN-complex (MRE11-RAD50-NBS1 complex), a prominent DNA strand resection and broken end bridging component involved in homologous recombination repair (HRR) and alternative non-homologous end joining (a-NHEJ). We analyzed the spatial arrangements of antibody-labelled MRE11 proteins in the nuclei of a breast cancer and a skin fibroblast cell line along a time-course of repair (up to 48 h) after irradiation with a dose of 2 Gy. Different kinetics for cluster formation and relaxation were determined. Changes in the internal nano-scaled structure of the clusters were quantified and compared between the two cell types. The results indicate a cell type-dependent DNA damage response concerning MRE11 recruitment and cluster formation. The MRE11 data were compared to H2AX phosphorylation detected by γH2AX molecule distribution. These data suggested modulations of MRE11 signal frequencies that were not directly correlated to DNA damage induction. The application of SMLM in radiation biophysics offers new possibilities to investigate spatial foci organization after DNA damaging and during subsequent repair. KW - Krebs, Medizin KW - Strahlentherapie KW - Brustkrebs Y1 - 2018 U6 - https://doi.org/https://doi.org/10.3390/cancers10010025 SN - 2072-6694 VL - 10 IS - 1 PB - MDPI AG ER - TY - JOUR A1 - Hausmann, Michael A1 - Winkler, Ralph A1 - Hildenbrand, Georg A1 - Finsterle, Jutta A1 - Weisel, Andrea A1 - Rapp, Alexander A1 - Schmitt, Eberhard A1 - Janz, Siegfried A1 - Cremer, Christoph T1 - COMBO-FISH: specific labeling of nondenatured chromatin targets by computer-selected DNA oligonucleotide probe combinations JF - BioTechniques KW - Genom Y1 - 2003 U6 - https://doi.org/https://doi.org/10.2144/03353rr03 SN - 0736-6205 VL - 35 IS - 3 SP - 564 EP - 577 PB - Informa UK Limited ER - TY - JOUR A1 - Wiech, Thorsten A1 - Stein, Stefan A1 - Lachenmaier, Victoria A1 - Schmitt, Eberhard A1 - Schwarz-Finsterle, Jutta A1 - Wiech, Elisabeth A1 - Hildenbrand, Georg A1 - Werner, Martin A1 - Hausmann, Michael T1 - Spatial allelic imbalance of BCL2 genes and chromosome 18 territories in nonneoplastic and neoplastic cervical squamous epithelium JF - European Biophysics Journal KW - Genom KW - Krebs, Medizin Y1 - 2009 U6 - https://doi.org/https://doi.org/10.1007/s00249-009-0474-5 SN - 0175-7571 VL - 38 IS - 6 SP - 793 EP - 806 PB - Springer Science and Business Media LLC ER -