TY - JOUR
T1 - Induction of AT-specific DNA-interstrand crosslinks by bizelesin in genomic and simian virus 40 DNA
AU - Woynarowski, Jan M.
AU - Chapman, William G.
AU - Napier, Cheryl
AU - Herzig, Maryanne C.S.
N1 - Funding Information:
The authors thank Dr. Laurence Hurley and the members of his research team for stimulating discussions and sharing their results prior to publication and Drs. R. Wadkins, D. Sun, and J. Quada for critical reading of the manuscript. This study was supported in part by a grant from the National Cancer Institute (CA71969) and by CTRC Research Foundation, San Antonio, TX.
PY - 1999/2/16
Y1 - 1999/2/16
N2 - Bizelesin is a bifunctional AT-specific DNA alkylating drug. Our study characterized the ability of bizelesin to induce interstrand crosslinks, a potential lethal lesion. In genomic DNA of BSC-1 cells, bizelesin formed from approx. 0.3 to 6.03±0.85 interstrand crosslinks per 106 base pairs, at 5- 100 nM drug concentration, respectively, comparable to the number of total adducts previously determined in the same system (J.M. Woynarowski, M.M. McHugh, L.S. Gawron, T.A. Beerman, Biochemistry 34 (1995) 13042-13050). Bizelesin did not induce DNA-protein crosslinks or strand breaks. A model defined target, intracellular simian virus 40 (SV40) DNA, was employed to map at the nucleotide level sites of bizelesin adducts, including potential interstrand crosslinks. Preferential adduct formation was observed at AT tracts which are abundant in the SV40 matrix associated region and the origin of replication. Many sites, including each occurrence of 5'-T(A/T)4A-3', co- mapped on both DNA strands suggesting interstrand crosslinks, although monoadducts were also formed. Bizelesin adducts in naked SV40 DNA were found at similar sites. The localization of bizelesin-induced crosslinks in AT- rich tracts of replication-related regions is consistent with the potent anti-replicative properties of bizelesin. Given the apparent lack of other types of lesions in genomic DNA, interstrand crosslinks localized in AT-rich tracts, and to some extent perhaps also monoadducts, are likely to be lethal effects of bizelesin.
AB - Bizelesin is a bifunctional AT-specific DNA alkylating drug. Our study characterized the ability of bizelesin to induce interstrand crosslinks, a potential lethal lesion. In genomic DNA of BSC-1 cells, bizelesin formed from approx. 0.3 to 6.03±0.85 interstrand crosslinks per 106 base pairs, at 5- 100 nM drug concentration, respectively, comparable to the number of total adducts previously determined in the same system (J.M. Woynarowski, M.M. McHugh, L.S. Gawron, T.A. Beerman, Biochemistry 34 (1995) 13042-13050). Bizelesin did not induce DNA-protein crosslinks or strand breaks. A model defined target, intracellular simian virus 40 (SV40) DNA, was employed to map at the nucleotide level sites of bizelesin adducts, including potential interstrand crosslinks. Preferential adduct formation was observed at AT tracts which are abundant in the SV40 matrix associated region and the origin of replication. Many sites, including each occurrence of 5'-T(A/T)4A-3', co- mapped on both DNA strands suggesting interstrand crosslinks, although monoadducts were also formed. Bizelesin adducts in naked SV40 DNA were found at similar sites. The localization of bizelesin-induced crosslinks in AT- rich tracts of replication-related regions is consistent with the potent anti-replicative properties of bizelesin. Given the apparent lack of other types of lesions in genomic DNA, interstrand crosslinks localized in AT-rich tracts, and to some extent perhaps also monoadducts, are likely to be lethal effects of bizelesin.
KW - AT-rich region
KW - Bizelesin
KW - DNA alkylation
KW - Interstrand crosslink
KW - Mapping DNA binding site
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U2 - 10.1016/S0167-4781(99)00002-0
DO - 10.1016/S0167-4781(99)00002-0
M3 - Article
C2 - 10023060
AN - SCOPUS:0033573947
VL - 1444
SP - 201
EP - 217
JO - Biochimica et Biophysica Acta - Gene Structure and Expression
JF - Biochimica et Biophysica Acta - Gene Structure and Expression
SN - 0167-4781
IS - 2
ER -