We examined DNA damage induced by the enediyne-containing antitumor an
tibiotic C-1027 in intracellular nuclear and mitochondrial DNA targets
using the episome-containing cell line 935.1. Strand-scission activit
y of the C-1027 holoantibiotic was measured by the topological forms c
onversion assay in episomal and mitochondrial DNA, as well as in cell-
free plasmid DNA. Genomic DNA damage was quantitated by filter elution
analysis. Comparisons were made to the well-characterized enediyne ne
ocarzinostatin. From these studies, mixed single- and double-strand br
eaks were observed not only in cell-free, plasmid DNA but also in intr
acellular episomal, mitochondrial, and genomic DNA at low nanomolar co
ncentrations. C-1027 cleaved DNA 285-fold more efficiently in cells th
an in a cell-free environment, and displayed preference for intracellu
lar DNA species in the following rank order: episome > mitochondrial D
NA >> genomic. NCS also damaged the non-histone-associated mitochondri
al DNA, but not the episome. Cleavage of the 935.1 cell episome by C-1
027 occurred at specific sites including the BPV origin of replication
and E6/E7 open reading frame regions, as well as the MMTV LTR promote
r region.