Nuclear foci of mammalian recombination proteins are located at single-stranded DNA regions formed after DNA damage.
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.
Max Planck Institute of Molecular Genetics, Ihnestrasse 73, 14195 Berlin, Germany.
E
E
Raderschall
E I
EI
Golub
T
T
Haaf
D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin / Q000502:physiology / Q000528:radiation effects; D003513:Cycloheximide / Q000494:pharmacology; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000032:analysis*; D003609:Dactinomycin / Q000494:pharmacology; D005047:Etoposide / Q000633:toxicity; D005347:Fibroblasts; D005720:Gamma Rays; D006801:Humans; D007700:Kinetics; D008322:Mammals; D008540:Meiosis; D016685:Mitomycin / Q000633:toxicity; D009687:Nuclear Proteins / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012680:Sensitivity and Specificity; D014983:Xeroderma Pigmentosum
D002843:Chromatin; D004277:DNA, Single-Stranded; D009687:Nuclear Proteins; D011994:Recombinant Proteins; D003609:Dactinomycin; D016685:Mitomycin; D005047:Etoposide; D003513:Cycloheximide