Unwinding and hydrodynamic flow linear dichroism characteristics of supercoiled DNA covalently modified with two isomeric methylchrysene diol epoxides of different biological activities.
Adducts derived from the covalent binding of two positional monomethyl-substituted isomers of a bay region chrysene diol epoxide to supercoiled pIBI30 DNA (2926 base pairs/genome) were prepared, and their characteristics were investigated by a combination of gel electrophoresis and flow linear dichroism techniques. The 5- and 6-methyl derivatives of trans-1,2-dihydroxy-anti-3,4-epoxy-1,2,3,4-tetrahydrochrysene [(+)-5- and (+)-6-MeCDE, respectively], both with 1R,2S,3S,4R stereochemistry, are characterized by significant differences in their biological activities [Melikian et al. (1988) Cancer Res. 48, 1781-1787]. When covalently bound to plasmid DNA, these two molecules give rise to striking differences in the gel electrophoretic and flow hydrodynamic characteristics of the modified supercoiled DNA. The hydrodynamic flow linear dichroism of linearized DNA molecules (obtained by EcoRI enzyme digestion of covalently closed supercoiled pIBI30 DNA), modified covalently with the highly tumorigenic and mutagenic (+)-5-MeCDE derivative, indicates that flexible joints, bends, or kinks are formed at the site of binding of (+)-5-MeCDE. Slab gel data, as well as ethidium bromide-titration tube agarose gel electrophoresis data, indicate that the formation of (+)-5-MeCDE-DNA lesions causes the removal of superhelical turns with an unwinding angle of 13 +/- 3 degrees per covalently bound polycyclic aromatic residue. In contrast, the biological inactive (+)-6-MeCDE does not significantly alter the characteristics of supercoiled DNA, the unwinding angle is only 2.7 +/- 1 degrees, and the changes in persistence lengths detected by the flow linear dichroism technique are significantly smaller than in the case of (+)-5-MeCDE-DNA adducts.(ABSTRACT TRUNCATED AT 250 WORDS)
Balasta
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Chemistry Department, New York University, New York 10003.
Xu
R
R
Geacintov
N E
NE
Swenberg
C E
CE
Amin
S
S
Hecht
S S
SS
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002911:Chrysenes / Q000737:chemistry*; D004269:DNA, Bacterial / Q000737:chemistry*; D004278:DNA, Superhelical / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D009532:Nickel / Q000737:chemistry; D009690:Nucleic Acid Conformation; D010957:Plasmids; D013237:Stereoisomerism
D002911:Chrysenes; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D009532:Nickel; C037783:1,2-dihydroxy-epoxy-1,2,3,4-tetrahydro-5-methylchrysene; C050280:1,2-dihydroxy-3,4-epoxy-1,2,3,4-tetrahydro-6-methylchrysene