beyond repair foci dna double-strand break repair in euchromatic and heterochromatic compartments analyzed by transmission electron microscopy无法修复病灶常染色质的dna双链断裂修复和异色的隔间通过透射电子显微镜分析.pdfVIP

Beyond Repair Foci: DNA Double-Strand Break Repair in Euchromatic and Heterochromatic Compartments Analyzed by Transmission Electron Microscopy 1. 1. 1 2 ¨ 1 Yvonne Lorat , Stefanie Schanz , Nadine Schuler , Gunther Wennemuth , Christian Rube , ¨ 1* Claudia E. Rube 1 Department of Radiation Oncology, Saarland University, Homburg/Saar, Germany, 2 Department of Anatomy and Cell Biology, Saarland University, Homburg/Saar, Germany Abstract Purpose: DNA double-strand breaks (DSBs) generated by ionizing radiation pose a serious threat to the preservation of genetic and epigenetic information. The known importance of local chromatin configuration in DSB repair raises the question of whether breaks in different chromatin environments are recognized and repaired by the same repair machinery and with similar efficiency. An essential step in DSB processing by non-homologous end joining is the high-affinity binding of Ku70-Ku80 and DNA-PKcs to double-stranded DNA ends that holds the ends in physical proximity for subsequent repair. Methods and Materials: Using transmission electron microscopy to localize gold-labeled pKu70 and pDNA-PKcs within nuclear ultrastructure, we monitored the formation and repair of actual DSBs within euchromatin (electron-lucent) and heterochromatin (electron-dense) in cortical neurons of irradiated mouse brain. Results: While DNA lesions in euchromatin (characterized by two pKu70-gold beads, reflecting the Ku70-Ku80 heterodimer) are promptly sensed and rejoined, DNA packaging in heterochromatin appears to retard DSB processing, due to the time needed to unravel higher-order chromatin structures. Complex pKu70-clusters formed