Characterization of Iron(II)·Bleomycin-Mediated RNA Strand Scission

Chris E. Holmes, Barbara J. Carter, Sidney M. Hecht

Research output: Contribution to journalArticlepeer-review

83 Scopus citations

Abstract

The ability of iron(II)·bleomycin to mediate RNA degradation was further characterized. At micromolar concentrations, FeII·BLM was shown to effect cleavage of Escherichia coli tRNA1His and a Schizosaccharomyces pombe amber suppressor tRNA construct in an efficient fashion. In contrast, E. coli tRNACys and yeast mitochondrial tRNAAsP and tRNAfMet precursors were not substrates for FeII·BLM. Also shown to be a good substrate for cleavage by FeII·BLM was yeast 5S ribosomal RNA. Since HIV-1 reverse transcriptase mRNA has previously been shown to be degraded by FeII·BLM (Carter et al., 1990a), members of the three major classes of RNA have now been shown to undergo Fe·BLM-mediated strand scission. For each of the substrate RNAs, cleavage occurred at sites unique to that substrate. Although RNA cleavage occurred at numerous sequences, 5′-G-pyr-3′ sites were prominent. Likewise, while cleavage was noted in regions anticipated to be double-stranded, as well as in single-stranded regions, a disproportionate number of cleavages were noted at the junction between single- and double-stranded regions. As found in earlier studies, RNA cleavage was much more selective than DNA cleavage. Further, when RNA cleavage was carried out in the presence of reagents such as Mg2+, spermidine, and NaCl, the selectivity of cleavage was further enhanced. The highly selective and efficient cleavage of a number of RNA molecules reinforces our earlier suggestion that RNA may constitute a therapeutically relevant target for bleomycin.

Original languageEnglish (US)
Pages (from-to)4293-4307
Number of pages15
JournalBiochemistry
Volume32
Issue number16
DOIs
StatePublished - Apr 1 1993
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

Fingerprint

Dive into the research topics of 'Characterization of Iron(II)·Bleomycin-Mediated RNA Strand Scission'. Together they form a unique fingerprint.

Cite this