Molecular analysis of mutations induced by the intercalating agent ellipticine at the hisD3052 allele of Salmonella typhimurium TA98.
We have used DNA colony hybridization, the polymerase chain reaction (PCR), and direct DNA sequencing to determine the mutations induced by the intercalating agent ellipticine in Salmonella typhimurium TA98 in the presence of S9. Of 400 ellipticine-induced revertants that were selected at a mutant yield that was ninefold over the background, 85.5% contained a GC or CG deletion within a common CGCGCGCG hotspot; this deletion occurred among 47% of the spontaneous revertants. In addition to this hotspot, the ellipticine spectrum contained two deletion warmspots that reside opposite each other in looped-out regions of a possible DNA secondary structure. Ellipticine and its metabolites likely revert Salmonella strain TA98 by forming DNA adducts that promote slippage-mismatches and by stabilizing these slipped mismatched sequences via intercalation. The involvement of these mechanisms, along with a likely role for DNA secondary structures and a possible role for DNA gyrase, may account for the site specificity exhibited by ellipticine in strain TA98.
DeMarini
D M
DM
Genetic Toxicology Division, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711.
Abu-Shakra
A
A
Gupta
R
R
Hendee
L J
LJ
Levine
J G
JG
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004252:DNA Mutational Analysis; D004611:Ellipticines / Q000737:chemistry / Q000633:toxicity*; D005609:Free Radicals; D007364:Intercalating Agents / Q000633:toxicity; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000235:genetics; D013329:Structure-Activity Relationship
D004611:Ellipticines; D005609:Free Radicals; D007364:Intercalating Agents; C034192:ellipticine; D004247:DNA