Rates and Consequences of Recombination between rRNA Operons

Abstract

ABSTRACT A mutant strain of Escherichia coli was created by inserting a cassette encoding sucrose sensitivity and neomycin resistance ( sacB-neo ) into the small-subunit rRNA-encoding gene rrs in the rrnB operon. During growth in a complex medium, the cassette was lost from the population, and a complete rrs gene was restored at a rate of 5 × 10 −9 per cell division. Repair of this lesion required flanking regions of DNA that were similar to the six remaining intact rRNA operons and reestablished the full complement of seven rRNA operons. The relative fitness of strains with restored rrnB operons was 1 to 2% higher than that of the mutant strain. The rrnB operon normally contains a spacer region between the 16S and 23S rRNA-encoding genes that is similar in length and tRNA gene content to the spacer in rrnC , - E , and - G . In 2 of the 14 strains in which rrnB was restored, the spacer region had the same length as the spacer region in rrnA , - D , and - H . The requirement for flanking regions of nearly identical DNA and the replication of the spacer region from other rRNA operons during the repair of rrnB suggest that the restoration was accomplished via gene conversion. The rate of gene conversion was 10-fold less than the fixation of point mutations in the same region of the chromosome but was apparently sufficient to homogenize the sequences of rRNA genes in E. coli . These findings are discussed in the context of a conceptual model describing the presence of sequence heterogeneity in coevolving rRNA genes.