title: Sequence specificity of DNA alkylation by the antitumor natural product leinamycin.
abstract: Reaction with thiol converts the antitumor natural product leinamycin to an episulfonium ion that alkylates the N(7)-position of guanine residues in double-stranded DNA. The sequence specificity for DNA alkylation by this structurally novel compound has not previously been examined. It is reported here that leinamycin shows significant (>10-fold) preferences for alkylation at the 5'-G in 5'-GG and 5'-GT sequences. The sequence preferences for activated leinamycin are significantly different from that observed for the structurally simple episulfonium ion generated from 2-chloroethyl ethyl sulfide. DNA alkylation by activated leinamycin is inhibited by addition of salt (100 mM NaClO(4)), although the degree of inhibition is somewhat less than that seen for 2-chloroethyl ethyl sulfide. This result suggests that electrostatic interactions between the activated leinamycin and the N(7)-position of guanine residues facilitate efficient DNA alkylation. However, the observed sequence preferences for DNA alkylation by activated leinamycin do not correlate strongly with calculated sequence-dependent variations in the molecular electrostatic potential at the N(7)-atom of guanine residues in duplex DNA. Thus, electrostatic interactions between activated leinamycin and DNA do not appear to be the primary determinant for sequence specificity. Rather, the results suggest that sequence-specific noncovalent interactions of leinamycin with the DNA double helix on the 3'-side of the alkylated guanine residue play a major role in determining the preferred alkylation sites. Consistent with the notion that noncovalent binding plays an important role in DNA alkylation by leinamycin, experiments with 2'-deoxyoligonucleotide substrates confirm that the natural product does not alkylate single-stranded DNA under conditions where duplex DNA is efficiently alkylated.
authors.affiliation: Departments of Chemistry and Biochemistry, University of Missouri-Columbia, Columbia, Missouri 65211, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Zang
authors.forename: Kent S
authors.initials: KS
authors.lastname: Gates
mesh_terms: D000478:Alkylation / Q000187:drug effects; D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000737:chemistry / Q000494:pharmacology*; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D004247:DNA / Q000187:drug effects / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D005399:Fishes; D006147:Guanine / Q000378:metabolism; D007769:Lactams; D018942:Macrolides / Q000737:chemistry / Q000494:pharmacology*; D008297:Male; D008969:Molecular Sequence Data; D009151:Mustard Gas / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D010472:Perchlorates / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D017670:Sodium Compounds / Q000494:pharmacology; D013094:Spermatozoa / Q000378:metabolism; D013379:Substrate Specificity; D013463:Sulfuric Acid Esters / Q000494:pharmacology; D013844:Thiazoles / Q000737:chemistry / Q000494:pharmacology*; D013871:Thiones / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D018906:Antineoplastic Agents, Alkylating; D004277:DNA, Single-Stranded; D007769:Lactams; D018942:Macrolides; D009841:Oligonucleotides; D010472:Perchlorates; D017670:Sodium Compounds; D013463:Sulfuric Acid Esters; D013844:Thiazoles; D013871:Thiones; C059000:leinamycin; D006147:Guanine; C031278:2-chloroethyl ethyl sulfide; D004247:DNA; C031068:sodium perchlorate; D010744:Phosphoric Monoester Hydrolases; C007482:dimethyl sulfate; D009151:Mustard Gas