New insights into calicheamicin-DNA interactions derived from a model nucleosome system.
Using the Xenopus borealis 5S RNA gene, we have identified several new features of the interaction of calicheamicin (CAL), an enediyne antitumor agent, with nucleosomal and naked DNA targets. CAL-mediated DNA damage was generally reduced by incorporation of the DNA into a nucleosome. However, in one instance, the frequency of DNA damage was enhanced in the nucleosome compared to naked DNA. This increase in CAL damage may result from bending-induced changes in the target site, while the association of histone proteins with DNA in the nucleosome may generally reduce the affinity of CAL for its targets by imposing dynamic constraints on the DNA, by altering target structure, or by steric hindrance. One implication of these observations is that new structural features created by incorporation of DNA into chromatin may produce 'hot spots' for CAL-mediated DNA damage not apparent in naked DNA studies. In a second set of experiments, the orientation of CAL at damage sites in naked 5S rDNA was determined. The results suggest that minor groove width per se is not a major determinant of CAL target selection. Our studies support the generality of an oligopurine recognition element, with the additional requirement that the purine tract is interrupted at the 3'-end by a pyrimidine(s). To account for these observations, we propose a model in which CAL recognizes the unique structural and dynamic features associated with the 3'-end of an oligopurine tract. Finally, we conclude that the dyad axis of pseudosymmetry of the 5S rRNA gene nucleosome cannot be determined with any degree of certainty. This places significant limitations on the interpretation of results from the study of drug-DNA interactions with reconstituted nucleosomes.
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Division of Toxicology, Massachusetts Institute of Technology, Cambridge 02139, USA.
Salzberg
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Dedon
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D000617:Aminoglycosides; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D000903:Antibiotics, Antineoplastic / Q000737:chemistry / Q000494:pharmacology*; D001483:Base Sequence; D001665:Binding Sites; D002645:Chickens; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage; D004275:DNA, Ribosomal / Q000187:drug effects; D053281:Enediynes; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000187:drug effects / Q000235:genetics*; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D013329:Structure-Activity Relationship; D014981:Xenopus
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D000903:Antibiotics, Antineoplastic; D004275:DNA, Ribosomal; D053281:Enediynes; D009707:Nucleosomes; D012341:RNA, Ribosomal, 5S; C055955:calicheamicin gamma(1)I; D004247:DNA