Interaction of calicheamicin gamma1(I) and its related carbohydrates with DNA-protein complexes.
We report studies of the contribution of DNA structure, holding the sequence constant, to the affinity of calicheamicin gamma(1)(I) and its aryltetrasaccharide moiety for DNA. We used polynucleotide chains as models of known protein-binding sequences [the catabolite activator protein (CAP) consensus sequence, AP-1 and cAMP response element (CRE) sites] in their free and protein-bound forms. The proteins were selected to provide examples in which the minor-groove binding site for the carbohydrate is (CAP) or is not (GCN4) covered by the protein. Additionally, peptides related to the GCN4 and CREB families, which have different bending effects on their DNA-binding sites, were used. We observe that proteins of the CREB class, which induce a tendency to bend toward the minor groove at the center of the site, inhibit drug-cleavage sites located at the center of the free AP-1 or CRE DNA sites. In the case of GCN4, which does not induce DNA bending, there is no effect on calicheamicin cleavage of the CRE site, but we observe a GCN4-induced rearrangement of the cutting pattern in the AP-1 site. This effect may arise from either a subtle local conformational rearrangement not accompanied by bending or a localized reduction in DNA flexibility. Whereas GCN4 binding is not inhibited by the calicheamicin aryltetrasaccharide, binding of CAP to its DNA target is significantly inhibited, and calicheamicin cutting of DNA at the center of the CAP-DNA complex site is strongly reduced by protein binding. This result probably reflects steric inhibition of drug binding by the protein.
Sissi
C
C
Department of Pharmaceutical Sciences, University of Padova, Via Marzolo 5, 35131 Padua, Italy.
Aiyar
J
J
Boyer
S
S
Depew
K
K
Danishefsky
S
S
Crothers
D M
DM
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D001667:Binding, Competitive; D004247:DNA / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D053281:Enediynes; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000494:pharmacology; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D011494:Protein Kinases / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins; D014312:Trisaccharides / Q000737:chemistry / Q000378:metabolism
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D004268:DNA-Binding Proteins; D053281:Enediynes; D005656:Fungal Proteins; D009844:Oligosaccharides; D011134:Polysaccharides; D029701:Saccharomyces cerevisiae Proteins; D014312:Trisaccharides; C121660:calicheamicin gamma(1)I aryltetrasaccharide; C121659:calicheamicin gamma(1)I aryltrisaccharide; C055955:calicheamicin gamma(1)I; D004247:DNA; D011494:Protein Kinases