In Situ Visualization of DNA Double-Strand Break Repair in Human Fibroblasts

• Published in: Science (American Association for the Advancement of Science) Vol. 280; no. 5363; pp. 590 - 592
• Main Authors: Nelms, Benjamin E., Maser, Richard S., MacKay, James F., Lagally, Max G., John H. J. Petrini
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for the Advancement of Science 24.04.1998; American Association for the Advancement of Science; The American Association for the Advancement of Science
• Subjects: Biological and medical sciences; Bromodeoxyuridine - immunology; Bromodeoxyuridine - metabolism; Cell Line; Cell lines; Cell nucleus; Cell Nucleus - metabolism; Cells; Deoxyribonucleic acid; DNA; DNA - metabolism; DNA - radiation effects; DNA Damage; DNA Repair; DNA-Binding Proteins - metabolism; Electrons; Fibroblasts; Fluorescein-5-isothiocyanate; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Humans; Irradiation; Masers; Microscopy, Confocal; Molecular and cellular biology; Molecular biology; Molecular genetics; Mutagenesis. Repair; Observations; Persistence; Rad51 Recombinase; Stripes; Visualization; X-rays; Human; Double strand break; Visualization; Ionizing irradiation; DNA; Lesion; Method; Repair; Fibroblast; X ray irradiation
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.280.5363.590

A method was developed to examine DNA repair within the intact cell. Ultrasoft x-rays were used to induce DNA double-strand breaks (DSBs) in defined subnuclear volumes of human fibroblasts and DNA repair was visualized at those sites. The DSBs remained in a fixed position during the initial stages of DNA repair, and the DSB repair protein hMre11 migrated to the sites of damage within 30 minutes. In contrast, hRad51, a human RecA homolog, did not localize at sites of DNA damage, a finding consistent with the distinct roles of these proteins in DNA repair.