ISSN:
1617-4623
Keywords:
Escherichia coli-ada −
;
Adapted
;
Mutations
;
Spectrum
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary The adaptive response is one of the major repair pathways in Escherichia coli that removes DNA alkylation damage. To investigate the role of the adaptive response in mutagenesis, the E. coli gpt forward mutation assay system was used to determine the mutation spectrum of N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) in MNNG-adapted and unadapted GP120 (wild-type) and unadapted PJ5 (ada-5) cells. We observed that 34/37 mutations in the unadapted GP120 cells, 38/40 mutations in the adapted GP120 cells, and 10/10 mutations in the PJ5 cells were GC → AT transitions. The remaining 3/37 mutations in the unadapted GP120 cells were large insertions. The remaining 2/40 mutations in the adapted GP120 cells were transversions with one a GC → CG and the other an AT → CG. A surrounding sequence specificity of mutagenesis was observed for the GC → AT transitions in both the unadapted (GP120 and PJ5) and adapted (GP120) cells, with 70% of the unadapted PJ5, 68% of the unadapted GP120, and 61% of the adapted GP120 mutations occurring at the middle G of the sequence 5′—GG(A or T)—3′. Both strains also displayed a statistically significant preference for mutagenesis at guanine bases in the non-transcribed strand. The overall distribution of mutated sites in the gpt gene in adapted and unadapted cells was similar, although the rate of mutations at certain sites appeared different. These minor differences could result from either non-uniform repair of alkylation damage at different sites on the DNA, or altered processing of the alkylated bases to mutations in the adapted state. However, these results indicate that there are no major differences in the types, surrounding sequence specificity, or distribution of mutations in the adapted GP120, unadapted GP120, or the ada − PJ5 cells.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00331159
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