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

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

Research output: Contribution to journalArticle

38 Citations (Scopus)

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

Fingerprint

CC 1065
Oligodeoxyribonucleotides
DNA
Adenine
Drugs
Fragment
Thymus
Breakage
DNA sequences
Antibiotics
Binding sites
DNA Sequence
Sequencing
Specificity
Minor
Gels
Molecules
B-Form DNA
Streptomyces
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Needham-vandevanter, D. R., Hurley, L., 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

Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex. / Needham-vandevanter, Donald R.; Hurley, Laurence; Reynolds, Vincent L.; Theriault, Nicole Y.; Krueger, Williams C.; Wierenga, Wendall.

In: Nucleic Acids Research, Vol. 12, No. 15, 10.08.1984, p. 6159-6168.

Research output: Contribution to journalArticle

Needham-vandevanter, DR, Hurley, L, Reynolds, VL, Theriault, NY, Krueger, WC & Wierenga, W 1984, 'Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex', Nucleic Acids Research, vol. 12, no. 15, pp. 6159-6168. https://doi.org/10.1093/nar/12.15.6159
Needham-vandevanter DR, Hurley L, Reynolds VL, Theriault NY, Krueger WC, Wierenga W. Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex. Nucleic Acids Research. 1984 Aug 10;12(15):6159-6168. https://doi.org/10.1093/nar/12.15.6159
Needham-vandevanter, Donald R. ; Hurley, Laurence ; Reynolds, Vincent L. ; Theriault, Nicole Y. ; Krueger, Williams C. ; Wierenga, Wendall. / Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex. In: Nucleic Acids Research. 1984 ; Vol. 12, No. 15. pp. 6159-6168.
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