Modulation of Neocarzinostatin-mediated DNA Double Strand Damage by Activating Thiol: Deuterium Isotope Effects

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1992 Feb 25

PMID 1531872

Authors

S E McAfee

G W Ashley

Affiliations

Soon will be listed here.

Abstract

The neocarzinostatin chromophore causes double-strand damage at AGC sequences on DNA by concomitant 1'-oxidation at C and 5'-oxidation at the T on the complementary strand. The extent of this damage is dependent upon the structure of the thiol used for activation. Deuterium isotope effects suggest that this dependence on thiol structure may be due to internal quenching of one radical site of the activated chromophore by the hydrogen atoms of the thiol sidechain. The 12-mer d[GCAAGCGCTTGC] is treated with the neocarzinostatin chromophore and either sodium thioglycolate or [2-2H2]-thioglycolate, and the distribution of strand breaks is determined by gel electrophoresis. Two isotope effects are noted: an overall sequence-independent effect in which deuterated thioglycolate increases total strand damage by a factor of 2, and a sequence-specific effect by which deuteration increases the proportion of alkali-sensitive strand damage at C6 by an additional factor of 1.5. Methyl thioglycolate shows essentially identical behavior to that of thioglycolate anion, ruling out electrostatic effects as major contributors to the effect of thiol structure on the mode of DNA damage observed. A model for NCSC action consistent with these results is discussed.

References

Riddles P, Blakeley R, ZERNER B . Ellman's reagent: 5,5'-dithiobis(2-nitrobenzoic acid)--a reexamination. Anal Biochem. 1979; 94(1):75-81. DOI: 10.1016/0003-2697(79)90792-9. View

Kappen L, Goldberg I, Liesch J . Identification of thymidine-5'-aldehyde at DNA strand breaks induced by neocarzinostatin chromophore. Proc Natl Acad Sci U S A. 1982; 79(3):744-8. PMC: 345828. DOI: 10.1073/pnas.79.3.744. View

Williamson J, Celander D . Rapid procedure for chemical sequencing of small oligonucleotides without ethanol precipitation. Nucleic Acids Res. 1990; 18(2):379. PMC: 330294. DOI: 10.1093/nar/18.2.379. View

Kappen L, Goldberg I . Identification of 2-deoxyribonolactone at the site of neocarzinostatin-induced cytosine release in the sequence d(AGC). Biochemistry. 1989; 28(3):1027-32. DOI: 10.1021/bi00429a016. View

Povirk L, Goldberg I . Endonuclease-resistant apyrimidinic sites formed by neocarzinostatin at cytosine residues in DNA: evidence for a possible role in mutagenesis. Proc Natl Acad Sci U S A. 1985; 82(10):3182-6. PMC: 397739. DOI: 10.1073/pnas.82.10.3182. View

Zein N, Sinha A, McGahren W, Ellestad G . Calicheamicin gamma 1I: an antitumor antibiotic that cleaves double-stranded DNA site specifically. Science. 1988; 240(4856):1198-201. DOI: 10.1126/science.3240341. View

Povirk L, Goldberg I . Covalent adducts of DNA and the nonprotein chromophore of neocarzinostatin contain a modified deoxyribose. Proc Natl Acad Sci U S A. 1982; 79(2):369-73. PMC: 345736. DOI: 10.1073/pnas.79.2.369. View

Hensens O, Dewey R, Liesch J, Napier M, Reamer R, Smith J . Neocarzinostatin chromophore: presence of a highly strained ether ring and its reaction with mercaptan and sodium borohydride. Biochem Biophys Res Commun. 1983; 113(2):538-47. DOI: 10.1016/0006-291x(83)91759-x. View

SAMY T, Hu J, Meienhofer J, Lazarus H, Johnson R . A facile method of purification of neocarzinostatin, an antitumor protein. J Natl Cancer Inst. 1977; 58(6):1765-8. DOI: 10.1093/jnci/58.6.1765. View

10.

Kappen L, Goldberg I, Frank B, Worth Jr L, Christner D, Kozarich J . Neocarzinostatin-induced hydrogen atom abstraction from C-4' and C-5' of the T residue at a d(GT) step in oligonucleotides: shuttling between deoxyribose attack sites based on isotope selection effects. Biochemistry. 1991; 30(8):2034-42. DOI: 10.1021/bi00222a005. View

11.

Meschwitz S, Goldberg I . 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. Proc Natl Acad Sci U S A. 1991; 88(8):3047-51. PMC: 51381. DOI: 10.1073/pnas.88.8.3047. View

12.

Dedon P, Goldberg I . Sequence-specific double-strand breakage of DNA by neocarzinostatin involves different chemical mechanisms within a staggered cleavage site. J Biol Chem. 1990; 265(25):14713-6. View

13.

Povirk L, Houlgrave C . 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. Biochemistry. 1988; 27(10):3850-7. DOI: 10.1021/bi00410a049. View

14.

Kappen L, Chen C, Goldberg I . Atypical abasic sites generated by neocarzinostatin at sequence-specific cytidylate residues in oligodeoxynucleotides. Biochemistry. 1988; 27(12):4331-40. DOI: 10.1021/bi00412a021. View

15.

Lee S, Thivierge J, Goldberg I . DNA microstructural requirements for neocarzinostatin chromophore-induced direct strand cleavage. Nucleic Acids Res. 1989; 17(14):5809-25. PMC: 318198. DOI: 10.1093/nar/17.14.5809. View

16.

Hawley R, Kiessling L, Schreiber S . Model of the interactions of calichemicin gamma 1 with a DNA fragment from pBR322. Proc Natl Acad Sci U S A. 1989; 86(4):1105-9. PMC: 286634. DOI: 10.1073/pnas.86.4.1105. View

17.

Chin D, Zeng C, Costello C, Goldberg I . Sites in the diyne-ene bicyclic core of neocarzinostatin chromophore responsible for hydrogen abstraction from DNA. Biochemistry. 1988; 27(21):8106-14. DOI: 10.1021/bi00421a020. View

18.

Takeshita M, Kappen L, Grollman A, Eisenberg M, Goldberg I . Strand scission of deoxyribonucleic acid by neocarzinostatin, auromomycin, and bleomycin: studies on base release and nucleotide sequence specificity. Biochemistry. 1981; 20(26):7599-606. DOI: 10.1021/bi00529a039. View

19.

Povirk L, Goldberg I . Neocarzinostatin chromophore-DNA adducts: evidence for a covalent linkage to the oxidized C-5' of deoxyribose. Nucleic Acids Res. 1982; 10(20):6255-64. PMC: 326915. DOI: 10.1093/nar/10.20.6255. View

20.

Kappen L, Goldberg I . Deoxyribonucleic acid damage by neocarzinostatin chromophore: strand breaks generated by selective oxidation of C-5' of deoxyribose. Biochemistry. 1983; 22(21):4872-8. DOI: 10.1021/bi00290a002. View