Revisiting classical Escherichia coli cell division mutants by whole-genome sequencing

TL;DR

This paper uses whole-genome sequencing to analyze old E. coli cell division mutants, revealing the impact of historical mutagenesis methods on their genomes.

Contribution

The study provides the first complete genome sequences of original E. coli cell division mutants and highlights the effects of mutagenesis techniques.

Findings

Mutants obtained with nitrosoguanidine had 100 to 400 mutations in their genomes.

Transducing target alleles reduced mutations but co-transduced nearby mutations.

Site-directed mutagenesis preserved the genomic background.

Abstract

Over 60 years ago, researchers started the genetic analysis of bacterial cell division by isolating conditional, temperature-sensitive mutants of essential Escherichia coli cell division genes. These early mutants were obtained by mutagenesis with chemical agents that introduced dozens to hundreds of mutations in the bacterial genomes. In this work, we present the complete genome sequences of six of these original mutants on ftsA, ftsZ and ftsQ genes, along with two of the strains used to generate them. The genomes of mutants obtained by exposure to nitrosoguanidine had 100 to 400 mutations. Transducing target alleles into a new strain effectively reduced the number of mutations, but those near the target gene were co-transduced with it. In contrast, a mutant generated by site-directed mutagenesis maintained the genomic background intact. The genomic analysis improves our understanding of these foundational strains, offering insights into the effects of historical mutagenesis techniques. These findings underscore the importance of genomic characterization in ensuring accurate interpretations of experimental results in microbiological research.

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