Comparison of the sequence selectivity of the DNA-alkylating pluramycin antitumour antibiotics DC92-B and hedamycin.
The sequence selectivity of DNA alkylation by the recently isolated pluramycin antitumour antibiotic DC92-B has been investigated using two methods: a piperidine-induced strand-breaking procedure and a Taq DNA polymerase/linear amplification method. These techniques reveal that guanines are the most reactive sites for alkylation and that the level of adduct formation at these sites is clearly sequence dependent. The highest levels of alkylation occurred at isolated guanines located in 5'-CGT sequences and also at the 5'-G in some 5'-CGG sequences. Isolated guanines in 5'-TGT sequences were also quite reactive. We have also re-examined, in parallel, the sequence selectivity of binding of the structurally-related compound hedamycin: the first known example of a bis(epoxide)-containing, DNA-alkylating pluramycin. Our studies included a more extensive sequence analysis of hedamycin binding than that previously reported and we are able, therefore, to define more precisely the sequence preference. Despite significant differences in the stereochemistry and substitution of their bis(epoxide) sidechains, hedamycin and DC92-B exhibited very similar sequence selectivities in our assays.
Prakash
A S
AS
Peter MacCallum Cancer Institute, Melbourne, Victoria, Australia.
Moore
A G
AG
Murray
V
V
Matias
C
C
McFadyen
W D
WD
Wickham
G
G
D000478:Alkylation; D000818:Animals; D000880:Anthraquinones / Q000378:metabolism* / Q000494:pharmacology*; D000903:Antibiotics, Antineoplastic / Q000378:metabolism* / Q000494:pharmacology*; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D004247:DNA / Q000187:drug effects* / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D008969:Molecular Sequence Data; D019384:Nucleic Acid Synthesis Inhibitors; D013053:Spectrophotometry; D019914:Taq Polymerase
D000880:Anthraquinones; D000903:Antibiotics, Antineoplastic; D019384:Nucleic Acid Synthesis Inhibitors; C001328:hedamycin; C057306:DC92-B; D004247:DNA; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI