Dynamics of bleomycin interaction with a strongly bound hairpin DNA substrate, and implications for cleavage of the bound DNA.
Recent studies involving DNAs bound strongly by bleomycins have documented that such DNAs are degraded by the antitumor antibiotic with characteristics different from those observed when studying the cleavage of randomly chosen DNAs in the presence of excess Fe·BLM. In the present study, surface plasmon resonance has been used to characterize the dynamics of BLM B(2) binding to a strongly bound hairpin DNA, to define the effects of Fe(3+), salt, and temperature on BLM-DNA interaction. One strong primary DNA binding site, and at least one much weaker site, were documented. In contrast, more than one strong cleavage site was found, an observation also made for two other hairpin DNAs. Evidence is presented for BLM equilibration between the stronger and weaker binding sites in a way that renders BLM unavailable to other, less strongly bound DNAs. Thus, enhanced binding to a given site does not necessarily result in increased DNA degradation at that site; i.e., for strongly bound DNAs, the facility of DNA cleavage must involve other parameters in addition to the intrinsic rate of C-4' H atom abstraction from DNA sugars.
Center for BioEnergetics, Biodesign Institute, and Department of Chemistry & Biochemistry, Arizona State University, Tempe, Arizona 85287, USA.
Trevor C
TC
Bozeman
Rupesh
R
Nanjunda
Chenhong
C
Tang
Yang
Y
Liu
Zachary J
ZJ
Segerman
Paul A
PA
Zaleski
W David
WD
Wilson
Sidney M
SM
Hecht
D000903:Antibiotics, Antineoplastic / Q000737:chemistry* / Q000494:pharmacology; D001761:Bleomycin / Q000737:chemistry* / Q000494:pharmacology; D004247:DNA / Q000737:chemistry* / Q000187:drug effects; D053837:DNA Cleavage / Q000187:drug effects; D015394:Molecular Structure; D020349:Surface Plasmon Resonance; D013816:Thermodynamics
D000903:Antibiotics, Antineoplastic; D001761:Bleomycin; D004247:DNA