The mutation that makes Escherichia coli resistant to λ P gene-mediated host lethality is located within the DNA initiator gene dnaA of the bacterium

Abstract

Earlier, we reported that the bacteriophage λ P gene product is lethal to Escherichia coli, and the E. coli rpl mutants are resistant to this λ P gene-mediated lethality. In this paper, we show that under the λ P gene-mediated lethal condition, the host DNA synthesis is inhibited at the initiation step. The rpl8 mutation maps around the 83 min position in the E. coli chromosome and is 94% linked with the dnaA gene. The rpl8 mutant gene has been cloned in a plasmid. This plasmid clone can protect the wild-type E. coli from λ P gene-mediated killing and complements E. coli dnaAts46 at 42° C. Also, starting with the wild-type dnaA gene in a plasmid, the rpl-like mutations have been isolated by in vitro mutagenesis. DNA sequencing data show that each of the rpl8, rpl12 and rpl14 mutations has changed a single base in the dnaA gene, which translates into the amino acid changes N313T, Y200N, and S246T respectively within the DnaA protein. These results have led us to conclude that the rpl mutations, which make E. coli resistant to λ P gene-mediated host lethality, are located within the DNA initiator gene dnaA of the host.