Gene- and tissue-specificity of mutation in Big Blue rats treated with the hepatocarcinogen N-hydroxy-2-acetylaminofluorene.
In a previous study, we found that treating transgenic Big Blue rats with the hepatocarcinogen N-hydroxy-2-acetylaminofluorene (N-OH-AAF) produced the same major DNA adduct in the target liver and the nontarget spleen lymphocytes and bone marrow cells, induced lacI mutants in the liver, and induced much lower frequencies of lacI and hprt mutants in spleen lymphocytes. In the present study, sequence analysis was conducted on lacI DNA and hprt cDNA from the mutants, to determine the mutational specificity of N-OH-AAF in the rat. All the mutation spectra from N-OH-AAF-treated rats differed significantly from corresponding mutation profiles from untreated animals (P = 0.02 to P < 0.0001). Although there were similarities among the mutational patterns derived from N-OH-AAF-treated rats (e.g., G:C --> T:A transversion was the most common mutation in all mutation sets), there were significant differences in the patterns of basepair substitution and frameshift mutation between the liver and spleen lymphocyte lacI mutants (P = 0.02) and between the spleen lymphocyte lacI and hprt mutants (P = 0.04). Also, multiplex PCR analysis of genomic DNA from the hprt mutants indicated that 12% of mutants from treated rats had major deletions in the hprt gene; no corresponding incidence of large deletions was evident among lacI mutations. All the mutation profiles reflect the general mutational specificity of the major DNA adduct formed by N-OH-AAF. The differences between N-OH-AAF mutation in the endogenous gene and transgene can be partially explained by the structures of the two genes. The tissue-specificity of the mutation spectra may contribute to targeting tumor formation to the liver. Environ. Mol. Mutagen. 37:203-214, 2001. Published 2001 Wiley-Liss, Inc.
Chen
T
T
Division of Genetic and Reproductive Toxicology, National Center for Toxicological Research, Jefferson, Arkansas 72079, USA. TChen@NCTR.FDA.GOV
Mittelstaedt
R A
RA
Shelton
S D
SD
Dass
S B
SB
Manjanatha
M G
MG
Casciano
D A
DA
Heflich
R H
RH
D000818:Animals; D030801:Animals, Genetically Modified; D001426:Bacterial Proteins / Q000187:drug effects / Q000235:genetics*; D001483:Base Sequence; D002273:Carcinogens / Q000633:toxicity*; D018736:DNA Adducts; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D029968:Escherichia coli Proteins; D005449:Fluorenes / Q000378:metabolism; D006881:Hydroxyacetylaminofluorene / Q000633:toxicity*; D007041:Hypoxanthine Phosphoribosyltransferase / Q000187:drug effects / Q000235:genetics*; D056968:Lac Repressors; D008099:Liver / Q000187:drug effects*; D008214:Lymphocytes / Q000187:drug effects; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D009928:Organ Specificity; D051381:Rats; D012097:Repressor Proteins / Q000187:drug effects / Q000235:genetics*; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013866:Thioguanine / Q000494:pharmacology
D001426:Bacterial Proteins; D002273:Carcinogens; D018736:DNA Adducts; D029968:Escherichia coli Proteins; D005449:Fluorenes; D056968:Lac Repressors; D012097:Repressor Proteins; C031838:N-(deoxyguanosin-8-yl)-2-aminofluorene; D006881:Hydroxyacetylaminofluorene; D007041:Hypoxanthine Phosphoribosyltransferase; D013866:Thioguanine; D003849:Deoxyguanosine