Biochemical basis of genotoxicity of heterocyclic arylamine food mutagens: Human DNA polymerase eta selectively produces a two-base deletion in copying the N2-guanyl adduct of 2-amino-3-methylimidazo[4,5-f]quinoline but not the C8 adduct at the NarI G3 site.
Heterocyclic arylamines are highly mutagenic and cause tumors in animal models. The mutagenicity is attributed to the C8- and N2-G adducts, the latter of which accumulates due to slower repair. The C8- and N 2-G adducts derived from 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) were placed at the G1 and G3 sites of the NarI sequence, in which the G3 site is an established hot spot for frameshift mutation with the model arylamine derivative 2-acetylaminofluorene but G1 is not. Human DNA polymerase (pol) eta extended primers beyond template G-IQ adducts better than did pol kappa and much better than pol iota or delta. In 1-base incorporation studies, pol eta inserted C and A, pol iota inserted T, and pol kappa inserted G. Steady-state kinetic parameters were measured for these dNTPs opposite the C8- and N 2-IQ adducts at both sites, being most favorable for pol eta. Mass spectrometry of pol eta extension products revealed a single major product in each of four cases; with the G1 and G3 C8-IQ adducts, incorporation was largely error-free. With the G3 N 2-IQ adduct, a -2 deletion occurred at the site of the adduct. With the G1 N 2-IQ adduct, the product was error-free at the site opposite the base and then stalled. Thus, the pol eta products yielded frame-shifts with the N 2 but not the C8 IQ adducts. We show a role for pol eta and the complexity of different chemical adducts of IQ, DNA position, and DNA polymerases.
Choi
Jeong-Yun
JY
Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232-0146, USA.
Stover
James S
JS
Angel
Karen C
KC
Chowdhury
Goutam
G
Rizzo
Carmelo J
CJ
Guengerich
F Peter
FP
D001671:Biochemistry / Q000379:methods; D004247:DNA / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry*; D017353:Gene Deletion; D006801:Humans; D007700:Kinetics; D008956:Models, Chemical; D011804:Quinolines / Q000737:chemistry*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000099270:Y-Family DNA Polymerases
D004247:DNA; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase; D015252:Deoxyribonucleases, Type II Site-Specific; D011804:Quinolines; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases; C046719:GGCGCC-specific type II deoxyribonucleases; C036989:2-amino-3,4-dimethylimidazo(4,5-f)quinoline