DNA cross-linking by intermediates in the mitomycin activation cascade.
We have assayed the cross-linking of oligonucleotides containing repeated mitomycin-reactive CpG sites in order to assess the factors that enhance activation of the carbamoyl function at C10, yielding efficient mitomycin cross-linking. Drugs studied include mitomycin C (MC), N-methylmitomycin A (NMA), and the aziridinomitosene of NMA (MS). Drugs were reduced both by catalytic hydrogenation and by diothionite. We find that cross-linking by fully reduced NMA can be increased severalfold by addition of either excess dithionite reductant or the oxidant FeCl3. Enhancement by FeCl3 is not seen with MC or MS, but excess dithionite increases cross-linking by all three compounds. We explain the action of Fe3+ by postulating production of the semiquinone of the monoadduct of mitomycin reacted at the C1-position; according to this mechanism, departure of the carbamate from C10 is more efficient for the semiquinone than for the hydroquinone. However, our results imply that the hydroquinone can also function as a cross-linking agent. Excess dithionite, beyond that required for stoichiometric reduction, activates the carbamate 2-3-fold for cross-linking. We find that the fully reduced leucoaziridinomitosene is highly unstable in solution, yet it produces efficient cross-liking. Hence, this compound is highly reactive in DNA alkylation and a good candidate for the role of primary alkylating agent.
Department of Chemistry, Yale University, New Haven, Connecticut 06511.
C
C
Cera
M
M
Egbertson
S P
SP
Teng
D M
DM
Crothers
S J
SJ
Danishefsky
D001483:Base Sequence; D055598:Chemical Phenomena; D002621:Chemistry; D003432:Cross-Linking Reagents; D004247:DNA; D007700:Kinetics; D008937:Mitomycins; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides
D003432:Cross-Linking Reagents; D008937:Mitomycins; D009838:Oligodeoxyribonucleotides; D004247:DNA