Characterization of an adduct between CC-1065 and a defined oligodeoxynucleotide duplex
Open Access
- 10 August 1984
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 12 (15) , 6159-6168
- https://doi.org/10.1093/nar/12.15.6159
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.Keywords
This publication has 5 references indexed in Scilit:
- An efficient method for the sequence analysis of oligodeoxyribonucleotidesAnalytical Biochemistry, 1983
- CC-1065 (NSC 298223), a novel antitumor agent that interacts strongly with double-stranded DNA.1982
- CC-1065 (NSC-298223), A MOST POTENT ANTI-TUMOR AGENT - KINETICS OF INHIBITION OF GROWTH, DNA-SYNTHESIS, AND CELL-SURVIVAL1982
- MECHANISM OF INTERACTION OF CC-1065 (NSC-298223) WITH DNA1982
- Mechanism of bleomycin action: studiesLife Sciences, 1981