N-Ethyl-N-nitrosourea induces A:T to C:G transversion mutations as well as transition mutations in SOS-induced Escherichiu coli

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Abstract
The fixation of DNA lesions induced in Escherichia coli by N-ethyl-N-nitrosourea (ENU) occurs by both SOS-dependent and SOS-independent pathways. To determine whether these pathways result in differential processing of ENU-induced lesions, we have analyzed the DNA sequence changes of mutations induced at a plasmidencoded herpes simplex virus type 1 thymidine kinase gene by ENU treatment of plasmidbearing RecA− and RecA+ bacteria, and by transformation of RecA−, RecA+ and SOS-induced RecA+ bacteria with ENU-modified plasmid DNA. Transition mutations were the predominant types of base substitution mutations observed for wild-type and RecA−E.coli, consistent with the SOS-independent mispairing of O6-ethylguanine and O4-ethylthymine adducts during DNA replication. Under conditions of SOS processing of ENU lesions, however, we observed the frequent induction of A:T – C:G transversion mutations. The proportion of A:T – C:G transversion mutations (42%) observed after transformation of SOS-induced bacteria with ENU modified DNA was approximately equal to that of the G:C – A:T transitions (46%). The frequencies of these mutations were increased 20- and 5-fold respectively over that observed for non-induced RecA+ cells. We suggest that ethylated DNA lesions which normally block DNA replication can be processed to yield A:T → C:G transversion mutations in SOS-induced E.coli.
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