Complex Frameshift Mutations Mediated by Plasmid pKM101: Mutational MechanismsDeduced from 4-Aminobiphenyl-Induced Mutation Spectra in Salmonella

摘要

The authors used colony probe hybridization and polymerase chain reaction/DNAsequence analysis to determine the mutations in approximately 2,400 4-aminobiphenyl (4-AB) + S9-induced revertants of the -1 frameshift allele hisD3052 and of the base-substitution allele hisG46 of Salmonella typhimurium. Most of the mutations occurred at sites containing guanine, which is the primary base at which 4-AB forms DNA adducts. A hotspot mutation involving the deletion of a CG or GC within the sequence CGCGCGCG accounted for 100 and 99.9%, respectively, of the reversion events at the hisD3052 allele in the pKM101 plasmid-minus strains TA1978 (uvr(+)) and TA1538 (Delta uvrB). In strain TA98 (DeltauvrB, pKM101), which contained the SOS DNA repair system provided by the pKM101 plasmid, approximately 85% of the revertants also contained the hotspot deletion; the remaining approximately 15% contained one of two types of mutations: (1) complex frameshifts that can be described as a -2 or +1 frameshift and an associated based substitution and (2) deletions of the CC or GG sequences that flank the hotspot site (CCGCGCGCGG). The authors propose a misincorporation/slippage model to account for these mutations in which (1) pKM101-mediated misincorporation and translesion synthesis occurs across a 4-AB-adducted guanine; (2) the instability of such a mispairing and/or the presence of the adduct leads to strand slippage in a run of repeated bases adjacent to the adducted guanine; and (3) continued DNA synthesis from the slipped intermediate produces a frameshift associated with a base substitution. (Copyright (c) 1994 by the Genetics Society of America.)