Binding of anti-Z-DNA antibodies to negatively supercoiled SV40 DNA

摘要

The binding of anti-Z-DNA antibody preparations to negatively supercoiled, protein-free SV40 DNA was analyzed. Covalent cross-linking with 0.1% glutaraldehyde followed by DNA restric tion endonucleolytic fragmentation and nitrocellulose filtration allowed accurate mapping of antibody binding sites. The critical superhelical density necessary to allow antibody binding was -a=0.056. The major region of antibody-DNA inter action was found within an SV40 segment spanning viral map positions 40 to 474. This region coincides with the nucleosome free region in SV40 minichromosomes and harbours the early and late promoter regions including the SV40 enhancer segment. Although it is unknown whether alternative, non-B-DNA confor mations are generated in vivo within SV40 minichromosomes our results emphasize the high degree of DNA structural flexibility that can be realized under negative torsional stress.