Selection-Enhanced Mutagenesis of lac Genes Is Due to Their Coamplification with dinB Encoding an Error-Prone DNA Polymerase

Genetics: A Periodical Record of Investigations Bearing on Heredity and Variation

article

Itsugo Yamayoshi¹  Sophie Maisnier-Patin¹  John R. Roth¹ 

[1] Department of Microbiology and Molecular Genetics, University of California, Davis, California 95616

关键词: dinB;    Escherichia coli;    adaptive mutation;    copy number variant;    error-prone polymerase;    gene amplification;    lactose operon;    local overreplication;    mutagenesis;    plasmid;   

DOI  :  10.1534/genetics.117.300409

学科分类：医学（综合）

来源: Genetics Society of America

【 摘 要 】

To test whether growth limitation induces mutations, Cairns and Foster constructed an Escherichia coli strain whose mutant lac allele provides 1–2% of normal ability to use lactose. This strain cannot grow on lactose, but produces ∼50 Lac⁺ revertant colonies per 10⁸ plated cells over 5 days. About 80% of revertants carry a stable lac⁺ mutation made by the error-prone DinB polymerase, which may be induced during growth limitation; 10% of Lac⁺ revertants are stable but form without DinB; and the remaining 10% grow by amplifying their mutant lac allele and are unstably Lac⁺. Induced DinB mutagenesis has been explained in two ways: (1) upregulation of dinB expression in nongrowing cells (“stress-induced mutagenesis”) or (2) selected local overreplication of the lac and dinB⁺ genes on lactose medium (selected amplification) in cells that are not dividing. Transcription of dinB is necessary but not sufficient for mutagenesis. Evidence is presented that DinB enhances reversion only when encoded somewhere on the F’ lac plasmid that carries the mutant lac gene. A new model will propose that rare preexisting cells (1 in a 1000) have ∼10 copies of the F’ lac plasmid, providing them with enough energy to divide, mate, and overreplicate their F’ lac plasmid under selective conditions. In these clones, repeated replication of F’ lac in nondividing cells directs opportunities for lac reversion and increases the copy number of the dinB⁺ gene. Amplification of dinB⁺ increases the error rate of replication and increases the number of lac⁺ revertants. Thus, reversion is enhanced in nondividing cells not by stress-induced mutagenesis, but by selected coamplification of the dinB and lac genes, both of which happen to lie on the F’ lac plasmid.

【 授权许可】

CC BY