The toxicity and bioactivity of pyrrolizidine alkaloids (PAs), common const
ituents of hundreds of plant species, and in herbal remedies and folk medic
ines prepared thereof, are probably due to their ability to form DNA cross-
linking. We investigated DNA cross-linking activity by chemically-activated
PAs from four different structural classes in Madin-Darby bovine kidney (M
DBK) cells and in pBR322 DNA. In cell culture, alpha,beta-unsaturated macro
cyclic diester pyrroles dehydrosenecionine (DHSN), dehydroriddelliine (DHRD
) and the saturated macrocyclic diester pyrrole dehydromonocrotaline (DHMO)
were significantly more potent cross-linkers than the simple necine base (
retronecine) and an N-oxide (indicine N-oxide; INO) as determined by alkali
ne elution. The proportion of total DNA cross-links that were proteinase K-
resistant (DNA-DNA cross-links) induced by the various pyrroles ranged from
0.08 (DHRN) to 0.67 (DHSN). Those pyrroles that were potent cross-linkers
of cellular DNA also cross-linked, in a dose-dependent manner, Bam Hi-diges
ted pBR322 DNA as assessed by a gel retardation assay. The possible functio
nal relevance of pyrrole-DNA cross-links was determined by their ability to
interrupt PCR amplification of a 1.129 kb segment of pBR322. Dehydroseneci
onine completely inhibited amplification, while DHMO was of intermediate po
tency, while DHRN and INO had no effect. Taken together, these studies sugg
est that structural features, most notably the presence of a macrocyclic di
ester, confer potent cross-link activity to PAs. In any event, DNA-DNA cros
s-linking is probably biologically relevant as indicated by their interfere
nce with DNA replication. (C) 1999 Elsevier Science Ltd. All rights reserve
d.