Correlation of frameshift mutagenicity with DNA intercalation by CGS 20928A using an in vitro DNA unwinding assay.
A compound's mutagenicity in different Salmonella tester strains can suggest its mechanism of reaction with DNA. Clear confirmation of such a mechanism, however, requires a direct test of the compound's reaction with DNA, often relying on specific in vitro studies. We report the use of a rapid in vitro test designed to measure DNA unwinding, a characteristic of DNA intercalators and many frameshift mutagens. CGS 20928A, an adenosine antagonist, produced a significant (> 2-fold) increase in revertants only for Salmonella tester strain TA1537, and only without metabolic activation. These data indicated that the compound was a direct acting frameshift mutagen and possibly intercalated into DNA. Our DNA unwinding assay indicated that at concentrations of > 0.1 mM CGS 20928A behaved like known intercalating compounds in that it unwound DNA. These concentrations of compound are comparable to those found mutagenic to TA1537. By comparison, the frameshift mutagen and known intercalating compound 9-aminoacridine unwound DNA in this assay in a concentration dependent fashion between 6-12 microM. ICR-191, another acridine frameshift mutagen, also unwound DNA. A compound structurally related to CGS 20928A, which was not mutagenic in Salmonella tester strains, did not produce any DNA unwinding even at 10 mM. Because the assay uses microgram quantities of material, it should be ideal for screening small amounts of congeneric series suspected of frameshift mutagenicity.
Mattes
W B
WB
Ciba, Farmington, Connecticut 06032.
Kapeghian
J C
JC
Lasinski
E R
ER
O'Lone
S D
SD
Puri
E C
EC
Matheson
D W
DW
D000241:Adenosine / Q000037:antagonists & inhibitors; D004269:DNA, Bacterial / Q000648:ultrastructure; D004270:DNA, Circular / Q000648:ultrastructure; D004305:Dose-Response Relationship, Drug; D016368:Frameshift Mutation; D005663:Furans / Q000737:chemistry / Q000633:toxicity*; D066298:In Vitro Techniques; D007364:Intercalating Agents; D015394:Molecular Structure; D009153:Mutagens; D009690:Nucleic Acid Conformation / Q000187:drug effects; D010957:Plasmids; D014230:Triazoles / Q000737:chemistry / Q000633:toxicity*
D004269:DNA, Bacterial; D004270:DNA, Circular; D005663:Furans; D007364:Intercalating Agents; D009153:Mutagens; D014230:Triazoles; C082400:CGS 20928A; D000241:Adenosine