A role for the metal binding domain in determining the DNA sequence selectivity of Fe-bleomycin

B. J. Carter, V. S. Murty, K. S. Reddy, S. N. Wang, S. M. Hecht

Research output: Contribution to journalArticle

95 Scopus citations

Abstract

Previous studies of Fe-bleomycin-mediated DNA cleavage have established that the bithiazole moiety + C-terminal substituent of bleomycin are required for DNA binding, while the metal binding domain is responsible for O2 activation. Although recent studies have indicated that the metal binding domain also participates in DNA unwinding, and in determining the sequence and strand selectivity of DNA cleavage, no study has defined the structural domain that bears primary responsibility for the observed pattern of bleomycin-mediated DNA degradation. Presently, by the use of four synthetic analogs of bleomycin demethyl A2 having the functional domains connected by rigid spacers of varying lengths, the source of DNA cleavage specificity has been determined. When the four analogs cleaved 242- and 127-base pair 5'-32P-end-labeled DNA restriction fragments containing isolated Fe-bleomycin cleavage sites, all four produced cleavage at the same preferred sites. Because the (oligo)glycine spacers altered the distance between the domains by as much as 14 Å, the identical cleavage patterns argue that the primary determinant of sequence specificity for these analogs is the metal binding domain.

Original languageEnglish (US)
Pages (from-to)4193-4196
Number of pages4
JournalJournal of Biological Chemistry
Volume265
Issue number8
StatePublished - Mar 30 1990
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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