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 complexes formed with calf thymus DNA. The above 14mer 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.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Banaszuk A. M., Deugau K. V., Sherwood J., Michalak M., Glick B. R. An efficient method for the sequence analysis of oligodeoxyribonucleotides. Anal Biochem. 1983 Feb 1;128(2):281–286. doi: 10.1016/0003-2697(83)90376-7. [DOI] [PubMed] [Google Scholar]
- Bhuyan B. K., Newell K. A., Crampton S. L., Von Hoff D. D. CC-1065 (NSC 298223), a most potent antitumor agent: kinetics of inhibition of growth, DNA synthesis, and cell survival. Cancer Res. 1982 Sep;42(9):3532–3537. [PubMed] [Google Scholar]
- Burger R. M., Peisach J., Horwitz S. B. Mechanism of bleomycin action: in vitro studies. Life Sci. 1981 Feb 16;28(7):715–727. doi: 10.1016/0024-3205(81)90153-3. [DOI] [PubMed] [Google Scholar]
- Li L. H., Swenson D. H., Schpok S. L., Kuentzel S. L., Dayton B. D., Krueger W. C. CC-1065 (NSC 298223), a novel antitumor agent that interacts strongly with double-stranded DNA. Cancer Res. 1982 Mar;42(3):999–1004. [PubMed] [Google Scholar]
- Swenson D. H., Li L. H., Hurley L. H., Rokem J. S., Petzold G. L., Dayton B. D., Wallace T. L., Lin A. H., Krueger W. C. Mechanism of interaction of CC-1065 (NSC 298223) with DNA. Cancer Res. 1982 Jul;42(7):2821–2828. [PubMed] [Google Scholar]