DNA methylation diminishes bleomycin-mediated strand scission

Robert P. Hertzberg, Mary Jo Caranfa, Sidney Hecht

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Three DNA duplexes differing substantially in sequence were derived from pBR322 plasmid DNA and supercoiled SV40 DNA by digestion with appropriate restriction endonucleases. Following treatment with the restriction methylase HhaI (recognition sequence: GCGC) or HhaI and HpaII (CCGG), the unmethylated and methylated DNAs were compared as substrates for the antitumor agent bleomycin. Bleomycin-mediated strand scission was shown to diminish substantially at a number of sites in proximity to the methylated cytidine moieties, especially where multiple sites had been methylated within a DNA segment of limited size. Detailed analysis of the DNA substrates revealed that both strands of DNA within a methylated region became more refractory to cleavage by bleomycin and that the protective effect could extend as many as 14 base pairs in proximity to the 5-methylcytidine moieties. Among the methylated DNA segments that became more resistant to bleomycin cleavage was a HpaII site of SV40 DNA, methylation of which has previously been shown to diminish the synthesis of the major late viral capsid protein following microinjection into Xenopus laevis oocytes. Study of the cleavage reaction at varying salt levels suggested that diminished bleomycin strand scission may be due, at least in part, to local conformational changes of the DNA to Z form (or other non-B-form structures). The results are generally consistent with the hypothesis that one mechanism for the expression of selective therapeutic action by certain DNA damaging agents could involve the recognition of specific methylation patterns.

Original languageEnglish (US)
Pages (from-to)5285-5289
Number of pages5
JournalBiochemistry
Volume24
Issue number20
StatePublished - 1985
Externally publishedYes

Fingerprint

Bleomycin
DNA Methylation
DNA
Z-Form DNA
Superhelical DNA
Cytidine
DNA Restriction Enzymes
Xenopus laevis
Capsid Proteins
Microinjections
Viral Proteins
Methylation
Base Pairing
Antineoplastic Agents
Substrates
Oocytes
Digestion
Refractory materials
Plasmids
Salts

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hertzberg, R. P., Caranfa, M. J., & Hecht, S. (1985). DNA methylation diminishes bleomycin-mediated strand scission. Biochemistry, 24(20), 5285-5289.

DNA methylation diminishes bleomycin-mediated strand scission. / Hertzberg, Robert P.; Caranfa, Mary Jo; Hecht, Sidney.

In: Biochemistry, Vol. 24, No. 20, 1985, p. 5285-5289.

Research output: Contribution to journalArticle

Hertzberg, RP, Caranfa, MJ & Hecht, S 1985, 'DNA methylation diminishes bleomycin-mediated strand scission', Biochemistry, vol. 24, no. 20, pp. 5285-5289.
Hertzberg, Robert P. ; Caranfa, Mary Jo ; Hecht, Sidney. / DNA methylation diminishes bleomycin-mediated strand scission. In: Biochemistry. 1985 ; Vol. 24, No. 20. pp. 5285-5289.
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