Further examination of the effects of nitrosylation on Alternaria alternata mycotoxin mutagenicity in vitro.
Previously, Alternaria extract and metabolite mutagenicities+/-nitrosylation were characterized using Ames Salmonella strains TA98 and TA100, which are both reverted at GC sites. To examine other targets for mutation, the metabolites Altertoxin I (ATX I), Altenuene (ALT), Alternariol (AOH), Alternariol monomethyl ether (AME), Tentoxin (TENT), Tenuazonic acid (TA) and Radicinin (RAD) were reexamined+/-nitrosylation, using Ames Salmonella strain TA97, sensitive to frameshift mutations at a run of C's, as well as strains TA102 and TA104, reverted by base pair mutations at AT sites and more sensitive to oxidative damage. ATX I was also assessed for mammalian mutagenicity at the Hprt gene locus in Chinese hamster V79 lung fibroblasts and rat hepatoma H4IIE cells. When tested from 1 to 100 microg/plate without nitrosylation, ATX I was mutagenic in TA102+/-rat liver S9 for activation and weakly mutagenic in TA104+/-S9, demonstrating direct-acting AT base pair mutagenicity. AOH was also directly mutagenic at AT sites in TA102+/-S9 while AME was weakly mutagenic in TA102+/-S9 and TA104+S9. Nitrosylation of ATX I enhanced mutagenicity at AT sites in TA104+/-S9 but produced little change in TA102+/-S9 compared to native ATX I. However, nitrosylated ATX I generated a potent direct-acting frameshift mutagen at C sites in TA97+/-S9. While ATX I was not mutagenic in either V79 cells or H4IIE cells, 5 and 10 microg/ml nitrosylated ATX I produced a doubling of 6-thioguanine resistant V79 colonies and 0.5 and 1 microg/ml were mutagenic to H4IIE cells, becoming toxic at higher concentrations. These results suggest ATX I, AME and AOH induce mutations at AT sites, possibly through oxidative damage, with nitrosylation enhancing ATX I frameshift mutagenicity at runs of C's. Nitrosylated ATX I was also directly mutagenic in mammalian test systems.
Schrader
T J
TJ
Toxicology Research Division, Bureau of Chemical Safety, Food Directorate, Health Canada, Ottawa, Ont., Canada K1A 0L2. tim_schrader@hc-sc.gc.ca
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D000528:Alternaria / Q000737:chemistry*; D000818:Animals; D006224:Cricetinae; D003412:Cricetulus; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008861:Microsomes / Q000378:metabolism; D009152:Mutagenicity Tests; D009153:Mutagens / Q000737:chemistry* / Q000494:pharmacology*; D009183:Mycotoxins / Q000737:chemistry* / Q000494:pharmacology*; D010569:Perylene / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics; D012977:Sodium Nitrite / Q000737:chemistry*
D009153:Mutagens; D009183:Mycotoxins; C050233:altertoxin I; D010569:Perylene; D007041:Hypoxanthine Phosphoribosyltransferase; D012977:Sodium Nitrite