Dependence of Z-DNA antibody binding to polytene chromosomes on acid fixation and DNA torsional strain.
There is considerable interest in the existence and significance of alternative conformations of DNA to the right-handed B-form described originally by Watson and Crick. The indirect immunofluorescence observations of Nordheim et al., Arndt-Jovin et al. and Lemeunier et al. that antibodies against left-handed Z-DNA bind to polytene chromosomes have thus assumed considerable importance. However, there is a paradox: some workers observe Z-DNA in interbands and others in bands. We report here that binding of Z-DNA antibodies to Drosophila polytene chromosomes prepared without acid fixation is at background level, and that following acid fixation the same antibody treatment leads to intense fluorescence. Depending on the extent of exposure to 45% acetic acid, fluorescence can occur primarily in interbands or in bands. Furthermore, antibody binding is dependent on elastic torsional strain in the DNA molecules.
D000818:Animals; D000936:Antigen-Antibody Complex; D002874:Chromosome Mapping / Q000379:methods*; D002875:Chromosomes / Q000648:ultrastructure*; D004247:DNA / Q000276:immunology*; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004278:DNA, Superhelical / Q000276:immunology; D004331:Drosophila melanogaster; D005404:Fixatives; D006863:Hydrogen-Ion Concentration
D000936:Antigen-Antibody Complex; D004278:DNA, Superhelical; D005404:Fixatives; D004247:DNA; D004264:DNA Topoisomerases, Type I