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

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 o...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 1998-04, Vol.280 (5363), p.590-592
Main Authors: Nelms, Benjamin E., Maser, Richard S., MacKay, James F., Lagally, Max G., John H. J. Petrini
Format: Article
Language:English
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
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Summary: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.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.280.5363.590