Identification of strong DNA binding motifs for bleomycin.
The bleomycins (BLMs) are clinically used antitumor antibiotics. Their mechanism of action is believed to involve oxidative cleavage of DNA and possibly also RNA degradation. DNA degradation has been studied extensively and shown to involve binding of an activated metallobleomycin to DNA, followed by abstraction of C4'-H from deoxyribose in the rate-limiting step for DNA degradation. It is interesting that while DNA and RNA degradation by activated Fe.BLM has been studied extensively, much less is known about the actual binding selectivity of BLM, that is, the obligatory step that precedes cleavage. Thus it is unclear whether cleavage specificity is defined by the binding event or whether cleavage occurs at a subset of preferred binding sites. With only a few exceptions, NMR binding studies have employed metalloBLMs such as Co.BLM and Zn.BLM whose therapeutic relevance is uncertain. A single biochemical study that compared DNA binding and cleavage directly also employed Co.BLM. It is logical to anticipate that preferred sites of DNA cleavage will occur at sites that are (a subset of) preferred DNA binding sites, but there are currently no data available relevant to this issue. Herein, we describe the development and implementation of a novel strategy to identify DNA motifs that bind BLM strongly.
Akiyama
Yoshitsugu
Y
Departments of Chemistry and Biology, University of Virginia, Charlottesville, Virginia 22904, USA.
Ma
Qian
Q
Edgar
Erin
E
Laikhter
Andrei
A
Hecht
Sidney M
SM
D000903:Antibiotics, Antineoplastic / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D001761:Bleomycin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D009690:Nucleic Acid Conformation; D013050:Spectrometry, Fluorescence
D000903:Antibiotics, Antineoplastic; D001761:Bleomycin; C025703:bleomycetin; D004247:DNA