Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex

Donald R. Needham-vandevanter, Laurence H. Hurley, Vincent L. Reynolds, Nicole Y. Theriault, Williams C. Krueger, Wendall Wierenga

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

CC-1065 is a potent antitumor antibiotic produced by Streptomyces zelensis. The drug binds covalently through N-3 of adenine and lies within the minor groove of DNA. Previous studies indicated that CC-1065 reacted with adenine in DNA to yield a thermally labile product that could be used to reveal its sequence specificity. These studies also provided insight into a DNA sequence (5′-CGGAGTTAGGGGCG-3′) which should bind one molecule of CC-1065 in an unambiguous manner. This sequence, which contains the CC-1065 adenine binding site within the sequence 5′-TTA-3′ was chemically synthesized together with the complementary strand. CC-1065 reacted with the oligoduplex to give an adduct that maintained the B-DNA form and had a final CD spectrum similar to those of the CC-1065 comp1exes formed with calf thymus DNA. The above l4mer was 5′ end-labelled with 32P, annealed with its complementary strand, reacted with CC-1065 and heated. Drug-mediated strand breakage was evaluated on a sequencing gel. A single break occurred in the labelled strands to give a fragment that migrated as an 8.5mer; subsequent piperidine treatment produced a fragment that migrated as a 7mer, which is the size expected from the known binding of CC-1065 at adenine in 5′-TTA-3′ sequences.

Original languageEnglish (US)
Pages (from-to)6159-6168
Number of pages10
JournalNucleic acids research
Volume12
Issue number15
DOIs
StatePublished - Aug 10 1984
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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    Needham-vandevanter, D. R., Hurley, L. H., Reynolds, V. L., Theriault, N. Y., Krueger, W. C., & Wierenga, W. (1984). Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex. Nucleic acids research, 12(15), 6159-6168. https://doi.org/10.1093/nar/12.15.6159