Inhibition of the BamHI cleavage and unwinding of pBR322 deoxyribonucleic acid by the antitumor drug cis-dichlorodiammineplatinum(II)

Henry Michael Ushay, T. D. Tullius, S. J. Lippard

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Abstract

The antitumour drug cis-dichlorodiammineplatinum(II) (cis-DDP) binds to pBR322 DNA and inhibits the cleavage of this circular DNA into a linear form by the restriction endonuclease BamHI. The binding of platinum to DNA was monitored by agarose gel electrophoresis, and the amount of platinum bound per nucleotide (r(b)) was measured by carbon rod atomic absorption spectroscopy. Electrophoretic mobility changes reflect a shortening and unwinding of the DNA duplex upon platinum binding as observed previously for the reaction of cis- and trans-DDP with pSM1 DNA. The inhibition of BamHI nuclease activity occurs at very low binding levels and is complete at r(b) = 0.045. This value corresponds to the binding of one platinum atom within 3 case pairs of the recognition sequence of the enzyme. Treatment of the DNA with 0.2 M sodium cyanide after BamHI cutting removes the platinum but does not alter the point at which cis-DDP inhibits the formation of the linear form III DNA. This result is in contrast with a previous report which claimed that BamHI could cut across a cis-DDP induced GpG cross-link in DNA which could be revealed subsequently by cyanide reversal of platinum binding. When the platinum is removed by cyanide treatment, the drug induced mobility changes are reversed and there is a pronounced sharpening of the bands in the gel. Quantitative study of the cyanide reversal shows the presence of a small amount of unremovable platinum tightly bound to the DNA at high ratios (0.1) of bound platinum per nucleotide.

Original languageEnglish (US)
Pages (from-to)3744-3748
Number of pages5
JournalBiochemistry
Volume20
Issue number13
StatePublished - 1981
Externally publishedYes

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Platinum
Antineoplastic Agents
Cisplatin
DNA
Cyanides
Deoxyribonuclease BamHI
Nucleotides
Gels
Sodium Cyanide
Circular DNA
Atomic spectroscopy
Electrophoretic mobility
DNA Cleavage
Agar Gel Electrophoresis
DNA Restriction Enzymes
Electrophoresis
Absorption spectroscopy
Sepharose
Spectrum Analysis
Carbon

ASJC Scopus subject areas

  • Biochemistry

Cite this

Inhibition of the BamHI cleavage and unwinding of pBR322 deoxyribonucleic acid by the antitumor drug cis-dichlorodiammineplatinum(II). / Ushay, Henry Michael; Tullius, T. D.; Lippard, S. J.

In: Biochemistry, Vol. 20, No. 13, 1981, p. 3744-3748.

Research output: Contribution to journalArticle

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