Publication Date:
2015-09-30
Description:
The roles of translesion synthesis (TLS) DNA polymerases in bypassing the C8–2'-deoxyguanosine adduct (dG-C8-IQ) formed by 2-amino-3-methylimidazo[4,5- f ]quinoline (IQ), a highly mutagenic and carcinogenic heterocyclic amine found in cooked meats, were investigated. Three plasmid vectors containing the dG-C8-IQ adduct at the G 1 -, G 2 - or G 3 -positions of the Nar I site (5'-G 1 G 2 CG 3 CC-3') were replicated in HEK293T cells. Fifty percent of the progeny from the G 3 construct were mutants, largely G-〉T, compared to 18% and 24% from the G 1 and G 2 constructs, respectively. Mutation frequency (MF) of dG-C8-IQ was reduced by 38–67% upon siRNA knockdown of pol , whereas it was increased by 10–24% in pol knockdown cells. When pol and pol were simultaneously knocked down, MF of the G 1 and G 3 constructs was reduced from 18% and 50%, respectively, to 〈3%, whereas it was reduced from 24% to 〈1% in the G 2 construct. In vitro TLS using yeast pol showed that it can extend G 3 *:A pair more efficiently than G 3 *:C pair, but it is inefficient at nucleotide incorporation opposite dG-C8-IQ. We conclude that pol and pol cooperatively carry out the majority of the error-prone TLS of dG-C8-IQ, whereas pol is involved primarily in its error-free bypass.
Print ISSN:
0305-1048
Electronic ISSN:
1362-4962
Topics:
Biology
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