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 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.
Nelms
B E
BE
Laboratory of Genetics and Department of Medical Physics, University of Wisconsin Medical School, Madison, WI 53706, USA.
Maser
R S
RS
MacKay
J F
JF
Lagally
M G
MG
Petrini
J H
JH
D001973:Bromodeoxyuridine / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D004247:DNA / Q000378:metabolism* / Q000528:radiation effects; D004249:DNA Damage; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism; D005347:Fibroblasts; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D006801:Humans; D018613:Microscopy, Confocal; D051135:Rad51 Recombinase
D004268:DNA-Binding Proteins; D004247:DNA; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; D001973:Bromodeoxyuridine; D016650:Fluorescein-5-isothiocyanate