title: Isolation of Drosophila proteins that bind selectively to left-handed Z-DNA.
abstract: An affinity column for isolating Z-DNA binding proteins was made by attaching brominated poly(dG-dC) to Sephadex. Proteins from Drosophila nuclei were prepared and those that could bind to Escherichia coli B-DNA were removed from the solution. The remaining proteins were passed over the Z-DNA affinity column and then eluted with NaCl. Using both direct and competitive filter binding assays, we found that the eluted proteins bind to brominated poly(dG-dC) (Z-DNA) and poly(dG-m5dC) but not to poly(dG-dC) (B-DNA), native or denatured E. coli or calf thymus DNA, or brominated oligonucleotides. The proteins also bind to negatively supercoiled plasmids carrying Z-DNA sequences but not to relaxed or linearized plasmids in which the Z-DNA conformation is no longer present. Gel analysis reveals a mixture of several large proteins up to approximately 150,000 daltons.
mesh_terms: D001667:Binding, Competitive; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism*; D004265:DNA Helicases / Q000302:isolation & purification*; D004278:DNA, Superhelical / Q000378:metabolism; D004268:DNA-Binding Proteins; D004331:Drosophila melanogaster / Q000032:analysis*; D008970:Molecular Weight; D009690:Nucleic Acid Conformation
chemical_list: D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D004247:DNA; D004265:DNA Helicases