Suppressor mutations (rin) that specifically suppress the recA+ dependence of stable DNA replication in Escherichia coliK-12

Abstract

The sdrA102 mutation confers upon cells the ability to replicate DNA in the absence of protein synthesis. This mutation was combined with the recA200 mutation, which renders the recA protein thermolabile, and had little effect on normal replication. However, the sdrA102 recA200 double mutant exhibited temperature-sensitive stable DNA replication: it replicated DNA continuously in the presence of chloramphenicol at 30 degrees C, whereas at 42 degrees C DNA replication ceased after the DNA content increased only 40-45%. Suppressor mutants (rin; recA-independent) capable of stable DNA replication at 42 degrees C were isolated from the double mutant. The suppressor mutant retained all other recA- characteristics, i.e., deficient general recombination, severe UV-sensitivity, and incapability of prophage induction in lysogens. This indicates that the rin mutation specifically suppresses the recA+ dependency of stable DNA replication. It is suggested that the recA+ protein stabilizes a specific structure, similar to an intermediate in recombination, which may function in the initiation of stable DNA replication.