C1027 chromophore, a potent new enediyne antitumor antibiotic, induces sequence-specific double-strand DNA cleavage.

C1027, a new macromolecular antitumor antibiotic produced by a Streptomyces strain, shows highly potent cytotoxicity to cultured cancer cells and marked DNA cleaving ability. The structure of its chromophore, responsible for most of the biological activities of the antibiotic, was recently determined and found to contain a nine-membered enediyne. In contrast to other enediyne antibiotics, such as neocarzinostatin, calicheamicin, esperamicin, and recently found kedarcidin, C1027 damages duplex DNA even in the absence of thiols. The DNA damage caused by C1027 includes double-strand breaks, single-strand breaks, and abasic sites. Experiments with plasmid DNA and 32P-end-labeled restriction fragments demonstrated that the chromophore, extracted from the protein-containing holoantibiotic, interacts in the DNA minor groove and cleaves double-helical DNA with a remarkable sequence-selectivity causing direct double-strand breaks. The double-strand cleavage sites, occurring predominantly at CTTTT/AAAAG, ATAAT/ATTAT, CTTTA/TAAAG, CTCTT/AAGAG, and especially GTTAT/ATAAC, consist of five nucleotide sequences with a two-nucleotide 3'-stagger of the cleaved residues (cutting sites are underlined). The chemical structures of the damaged residues at the GTTAT/ATAAC cleavage site suggest a model in which a C1027-induced double-strand break results from abstraction, by a single molecule of the diradical form of the chromophore, of a C4' hydrogen atom from the A residue of GTTAT and a C5' hydrogen atom from the A of ATAAC on the opposite strand. Single-strand breaks, which are mainly produced at adenylate and thymidylate residues, appear to be separate events presumably resulting from different binding modes of the drug to DNA.(ABSTRACT TRUNCATED AT 250 WORDS)

[1]  O. Hensens,et al.  Spontaneous generation of a biradical species of neocarzinostatin chromophore: role in DNA bulge-specific cleavage. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[2]  I. Goldberg,et al.  Site-specific cleavage at a DNA bulge by neocarzinostatin chromophore via a novel mechanism. , 1993, Biochemistry.

[3]  I. Goldberg,et al.  DNA conformation-induced activation of an enediyne for site-specific cleavage. , 1993, Science.

[4]  Stella Huang,et al.  Chemistry and structure elucidation of the kedarcidin chromophore , 1993 .

[5]  T. Ishii,et al.  Synthesis and absolute stereochemistry of the aminosugar moiety of antibiotic C-1027 chromophore , 1993 .

[6]  Y. Sugiura,et al.  Some characteristics of DNA strand scission by macromolecular antitumor antibiotic C-1027 containing a novel enediyne chromophore. , 1993, Biochemistry.

[7]  T. Otani Conformation studies on and assessment by spectral analysis of the protein-chromophore interaction of the macromolecular antitumor antibiotic C-1027. , 1993, The Journal of antibiotics.

[8]  Ken‐ichiro Yoshida,et al.  Structure of an aromatization product of C-1027 chromophore , 1993 .

[9]  T. Tullius,et al.  How the structure of an adenine tract depends on sequence context: a new model for the structure of TnAn DNA sequences. , 1993, Biochemistry.

[10]  C. Townsend,et al.  Specific abstraction of the 5'S- and 4'-deoxyribosyl hydrogen atoms from DNA by calicheamicin .gamma.1I , 1992 .

[11]  I. Goldberg,et al.  Neocarzinostatin acts as a sensitive probe of DNA microheterogeneity: switching of chemistry from C-1' to C-4' by a G.T mismatch 5' to the site of DNA damage. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[12]  P. Dedon,et al.  Free-radical mechanisms involved in the formation of sequence-dependent bistranded DNA lesions by the antitumor antibiotics bleomycin, neocarzinostatin, and calicheamicin. , 1992, Chemical research in toxicology.

[13]  P. Dedon,et al.  Neocarzinostatin-mediated DNA damage in a model AGT.ACT site: mechanistic studies of thiol-sensitive partitioning of C4' DNA damage products. , 1992, Biochemistry.

[14]  M. Lee,et al.  Calicheamicins: Discovery, Structure, Chemistry, and Interaction with DNA , 1991 .

[15]  I. Goldberg Mechanism of Neocarzinostatin Action: Role of DNA Microstructure in Determination of Chemistry of Bistranded Oxidative Damage , 1991 .

[16]  T. Otani,et al.  Isolation and characterization of non-protein chromophore and its degradation product from antibiotic C-1027. , 1991, The Journal of antibiotics.

[17]  S. Meschwitz,et al.  Selective abstraction of 2H from C-5' of thymidylate in an oligodeoxynucleotide by the radical center at C-6 of the diradical species of neocarzinostatin: chemical evidence for the structure of the activated drug-DNA complex. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[18]  P. Dedon,et al.  Sequence-specific double-strand breakage of DNA by neocarzinostatin involves different chemical mechanisms within a staggered cleavage site. , 1990, The Journal of biological chemistry.

[19]  Y. Sugiura,et al.  DNA intercalation and cleavage of an antitumor antibiotic dynemicin that contains anthracycline and enediyne cores. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Y. Sugimoto,et al.  Mechanism of action of a new macromolecular antitumor antibiotic, C-1027. , 1990, The Journal of antibiotics.

[21]  D. Vanderwall,et al.  Sequence-specific isotope effects on the cleavage of DNA by bleomycin. , 1989, Science.

[22]  H. Saito,et al.  A new macromolecular antitumor antibiotic, C-1027. III. Antitumor activity. , 1989, The Journal of antibiotics.

[23]  Y. Yamada,et al.  A new macromolecular antitumor antibiotic, C-1027. I. Discovery, taxonomy of producing organism, fermentation and biological activity. , 1988, The Journal of antibiotics.

[24]  T. Otani,et al.  A new macromolecular antitumor antibiotic, C-1027. II. Isolation and physico-chemical properties. , 1988, The Journal of antibiotics.

[25]  G. Ellestad,et al.  Calicheamicin gamma 1I: an antitumor antibiotic that cleaves double-stranded DNA site specifically. , 1988, Science.

[26]  L. Povirk,et al.  Effect of apurinic/apyrimidinic endonucleases and polyamines on DNA treated with bleomycin and neocarzinostatin: specific formation and cleavage of closely opposed lesions in complementary strands. , 1988, Biochemistry.

[27]  J. Stubbe,et al.  Mechanisms of bleomycin-induced DNA degradation , 1987 .

[28]  I. Goldberg,et al.  Deoxyribonucleic acid damage by neocarzinostatin chromophore: strand breaks generated by selective oxidation of C-5' of deoxyribose. , 1983, Biochemistry.

[29]  C. Iden,et al.  Bleomycin-induced strand-scission of DNA. Mechanism of deoxyribose cleavage. , 1981, The Journal of biological chemistry.

[30]  T. Hatayama,et al.  DNA damage and repair in relation to cell killing in neocarzinostatin-treated HeLa cells. , 1979, Biochimica et biophysica acta.

[31]  R. Lloyd,et al.  Bleomycin-specific fragmentation of double-stranded DNA. , 1978, Biochemistry.

[32]  L. Povirk,et al.  DNA double-strand breaks and alkali-labile bonds produced by bleomycin. , 1977, Nucleic acids research.

[33]  S. Schreiber,et al.  Site-specific atom transfer from DNA to a bound ligand defines the geometry of a DNA-calicheamicin .gamma.1I complex , 1990 .

[34]  J. Catino,et al.  Esperamicins, a class of potent antitumor antibiotics: mechanism of action. , 1989, Proceedings of the National Academy of Sciences of the United States of America.