On the Chemical Nature of DNA and RNA Modification by a Hemin Model System

Reuel B. Van Atta, Jean Bernadou, Bernard Meunier, Sidney M. Hecht

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

In order to model the interaction of hemin with DNA and other polynucleotides, we have studied the degradation of DNA, RNA, and polynucleotides of defined structure by [meso-tetrakis(N-methyl-4-pyridyl)porphinato]manganese(III) (MnTMPP) + KHSO5. The activated porphyrin was shown to release adenine, thymine, and cytosine from DNA; RNA degradation afforded adenine, uracil, and cytosine. The same products were obtained from single- and double-stranded DNA oligonucleotides of defined sequence, and also from single-stranded DNA and RNA homopolymers. The overall yield of bases from the dode-canucleotide d(CGCT3A3GCG) was equal to 14% of the nucleotides present initially, indicating that each porphyrin catalyzed the release of ~4 bases. Although no guanine was detected as a product from any of the substrates studied, the ability of MnTMPP + KHS05 to degrade guanine nucleotides was verified by the destruction of pGp, and by the appearance of bands corresponding to guanosine cleavage following treatment of 32P end labeled DNA restriction fragments with activated MnTMPP. Inspection of a number of sites of MnTMPP-promoted cleavage indicated that the process was sequence-selective, occurring primarily at G residues that were part of 5′-TG-3′ or 5′-AG-3′ sequences, or at T residues. Also formed in much greater abundance were alkali-labile lesions; these were formed largely at guanosine residues. Also studied was the degradation of a 47-nucleotide RNA molecule containing two hairpins. Degradation of the 5′-32P end labeled RNA substrate afforded no distinct, individual bands, suggesting that multiple modes of degradation may be operative. However, at concentrations of MnTMPP + KHSO5 that led to only limited amounts of RNA substrate degradation, there was enhanced degradation in a single-stranded region between the two hairpins, suggesting that MnTMPP may be a useful probe of RNA conformation.

Original languageEnglish (US)
Pages (from-to)4783-4789
Number of pages7
JournalBiochemistry
Volume29
Issue number20
DOIs
StatePublished - May 1 1990

ASJC Scopus subject areas

  • Biochemistry

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