Preferential replication-dependent mutagenesis in the lagging DNA strand in Escherichia coli.
The mutation frequencies attributable to -1 frameshift or one-base substitution in the structural genes coding for resistance to chloramphenicol (Cm) and tetracycline (Tc) were followed over several cycles of DNA replication, and found to differ several-fold, depending on the orientation of the gene on the plasmid with respect to the direction of (unidirectional ColE1-type) replication. The mutation frequency was higher when the reporter gene was present in the plasmid in the same orientation as the direction of the origin, i.e., when the transcription template is the lagging daughter strand, than when the gene was inserted in the opposite orientation. This significant difference in reversion frequencies of genes with different polarities was demonstrated only for a brief period of cell growth (several cycles of replication) after induction of the dnaQ49 mutator, but was not observed when an increased number of replication cycles, was permitted, most probably due to fixation of the mutation into both strands. The mutated intermediate DNA which possesses a misaligned basepair in the Cm gene was demonstrated to be replicated into two progeny DNA molecules; one is the chloramphenicol-resistant (CmR) DNA synthesized from the template strand having the mutation and the other is the CmS DNA from the template strand without mutation. Our results suggest that replication-dependent mutagenesis may occur preferentially in the lagging strand.
Iwaki
T
T
Department of Molecular Genetics, Kyoto Pharmaceutical University, Japan.
Kawamura
A
A
Ishino
Y
Y
Kohno
K
K
Kano
Y
Y
Goshima
N
N
Yara
M
M
Furusawa
M
M
Doi
H
H
Imamoto
F
F
D003086:Bacteriocin Plasmids / Q000235:genetics; D001483:Base Sequence; D002702:Chloramphenicol Resistance / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D016368:Frameshift Mutation / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics*; D017354:Point Mutation / Q000235:genetics; D013753:Tetracycline Resistance / Q000235:genetics
D004269:DNA, Bacterial