Replication and amplification of λ plasmids in Escherichia coli during amino acid starvation and limitation

Abstract

It was demonstrated previously that replication of plasmids derived from bacteriophage λ (so-called λ plasmids) is inhibited in wild-type Escherichia coli cells starved for isoleucine and arginine whereas it proceeds under the same conditions in relA mutants. Since replication of other replicons during the stringent or relaxed response depends on the nature of the deprived amino acid, we investigated replication of λ plasmids in E. coli relA + and relA − strains starved for different amino acids. We found that replication of λ plasmids is generally inhibited during the stringent, but not relaxed, response. Differences between cells starved for different amino acids, although reproducible, were not dramatic. Amino acid starvation was previously proposed as a method for amplification of λ plasmid DNA in vivo. We found that during amino acid limitation λ plasmids replicate more extensively in the relA mutants than during amino acid starvation. The efficiency of plasmid DNA amplification was found to be dependent on the kind of limited amino acid; in relA − bacteria limited for leucine we observed about 10-fold plasmid amplification. Some λ plasmid replication was also found under these conditions in the relA + host. The mechanism of the stringent control of λ plasmid DNA replication has already been proposed. Here the possible mechanism of the regulation of λ plasmid replication during amino acid limitation is presented.