Polκ protects mammalian cells against the lethal and mutagenic effects of benzo[a]pyrene

Abstract

Several low-fidelity DNA polymerases have recently been discovered that are able to bypass DNA lesions during DNA synthesis in vitro. The efficiency and accuracy of lesion bypass is, however, both polymerase and lesion specific. For example, in vitro studies revealed that human DNA polymerase κ (Polκ) is unable to insert a base opposite a cis-syn thymine-thymine dimer or cisplatin adduct, yet can bypass some DNA lesions such as a basic site and acetylaminofluorene-adducted guanine in an error-prone manner. More importantly, Polκ is able to bypass benzo[a]pyrene (B[a]P)-adducted guanine accurately and efficiently. To investigate the biological function of Polκ, we have generated mouse embryonic stem (ES) cells deficient in the Polκ gene encoding the enzyme. Polκ-deficient ES cells grow normally and their sensitivities to UV and x-ray radiation are only slightly affected. In contrast, the mutant cells are highly sensitive to both killing and mutagenesis induced by B[a]P. Furthermore, the spectrum of mutations recovered in the Polκ-deficient cells is different from that in the wild-type cells. Thus, our results indicate that Polκ plays an important role in suppressing mutations at DNA lesions generated by B[a]P.