Nucleic Acids Research, Vol 24, Issue 20 3918-3925, Copyright © 1996 by Oxford University Press
O Vrana, V Boudny and V Brabec
Negatively supercoiled, relaxed and linearized forms of pSP73 DNA were
modified in cell-free medium by cis-diamminedichloroplatinum(II)
(cisplatin). The frequency of interstrand cross-links (ICLs) formed in
these DNAs has been determined by: (i) immunochemical analysis; (ii) an
assay employing NaCN as a probe of DNA ICLs of cisplatin; (iii) gel
electrophoresis under denaturing conditions. At low levels of the
modification of DNA (<1 Pt atom fixed per 500 bp) the number of ICLs
formed by cisplatin was radically enhanced in supercoiled in comparison
with linearized or relaxed DNA. At these low levels of modification, the
frequency of ICLs in supercoiled DNA was enhanced with increasing level of
negative supercoiling or with decreasing level of modification. In
addition, the replication mapping of DNA ICLs of cisplatin was consistent
with these lesions being preferentially formed in negatively supercoiled
DNA between guanine residues in both the 5'- d(GC)-3' and the 5'-d(CG)-3'
sites. Among the DNA adducts of cisplatin the ICL has the markedly greatest
capability to unwind the double helix. We suggest that the formation of
ICLs of cisplatin is thermodynamically more favored in negatively
supercoiled DNA owing mainly to the relaxation of supercoils.
ARTICLES
Superhelical torsion controls DNA interstrand cross-linking by antitumor cis- diamminedichloroplatinum(II)
Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno.
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