Lupus-inducing drugs alter the structure of supercoiled circular DNA domains.
We analyzed the effects of procainamide (PROC), hydralazine (HYD), N-acetylprocainamide (NAPA), and L-canavanine (CAN) on circular supercoiled plasmids as models for chromosomal loop domains. The supercoil-dependent B-Z equilibrium in recombinant plasmids was used as an indicator of structural changes induced in circular DNA. Two-dimensional gel electrophoresis showed that PROC and HYD strongly inhibited supercoil-induced Z-DNA formation, whereas NAPA caused less pronounced changes in the B-Z equilibrium, and CAN had no effect. Gel retardation assays showed that the binding of a Z-DNA-specific autoimmune antibody to a Z-DNA-containing plasmid was strongly perturbed by HYD, but not influenced by CAN. Both PROC and NAPA showed moderate inhibition of antibody binding. Our results demonstrate the different potentials of these 4 drugs to interact with DNA and to alter the tertiary topology of DNA domains. It is conceivable that the in vivo capacity of PROC and HYD to induce antinuclear antibodies may be related to their ability to influence structural features in chromosomal DNA domains or nucleosomes, thus liberating antigenic structural epitopes in DNA and/or DNA-associated proteins.
Department of Medicine, University of Alabama, Birmingham, AL 35294.
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Zacharias
W J
WJ
Koopman
D000120:Acecainide / Q000494:pharmacology*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002172:Canavanine / Q000494:pharmacology*; D004278:DNA, Superhelical / Q000187:drug effects* / Q000276:immunology / Q000648:ultrastructure; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli; D006830:Hydralazine / Q000494:pharmacology*; D008180:Lupus Erythematosus, Systemic / Q000139:chemically induced*; D010957:Plasmids; D011342:Procainamide / Q000031:analogs & derivatives* / Q000494:pharmacology*
D000906:Antibodies; D004278:DNA, Superhelical; D006830:Hydralazine; D002172:Canavanine; D000120:Acecainide; D011342:Procainamide