Preferential replication dependent mutagenesis in the lagging DNA strand in Escherichia coli

Toshio Iwaki, Akira Kawamura, Yoshizumi Ishino, Kyoko Kohno, Yasunobu Kano, Naoki Goshima, Masaki Yara, Mitsuru Furusawa, Hirofumi Doi, Fumio Imamoto

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)

Abstract

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 (Cm(R)) DNA synthesized from the template strand having the mutation and the other is the Cm(S) DNA from the template strand without mutation. Our results suggest that replication-dependent mutagenesis may occur preferentially in the lagging strand.

Original languageEnglish
Pages (from-to)657-664
Number of pages8
JournalMolecular and General Genetics
Volume251
Issue number6
DOIs
Publication statusPublished - Oct 17 1996
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Genetics

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