Abstract
A sensitive and rapid in situ method was developed to visualize sites of single-stranded (ss) DNA in cultured cells and in experimental test animals. Anti-bromodeoxyuridine antibody recognizes the halogenated base analog incorporated into chromosomal DNA only when substituted DNA is in the single strand form. After treatment of cells with DNA-damaging agents or gamma irradiation, ssDNA molecules form nuclear foci in a dose-dependent manner within 60 min. The mammalian recombination protein Rad51 and the replication protein A then accumulate at sites of ssDNA and form foci, suggesting that these are sites of recombinational DNA repair.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Cell Cycle*
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Cell Line
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Cell Nucleus / metabolism*
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Chromatin / physiology
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Chromatin / radiation effects
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Cycloheximide / pharmacology
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DNA Damage*
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DNA, Single-Stranded / analysis*
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Dactinomycin / pharmacology
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Etoposide / toxicity
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Fibroblasts
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Gamma Rays
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Humans
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Kinetics
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Mammals
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Meiosis
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Mitomycin / toxicity
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Nuclear Proteins / drug effects
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Nuclear Proteins / metabolism*
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Nuclear Proteins / radiation effects
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Rats
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Recombinant Proteins / metabolism
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Sensitivity and Specificity
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Xeroderma Pigmentosum
Substances
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Chromatin
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DNA, Single-Stranded
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Nuclear Proteins
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Recombinant Proteins
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Dactinomycin
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Mitomycin
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Etoposide
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Cycloheximide