High levels of manganese-containing superoxide dismutase and thermally induced DNA disruption in a dnaK7(Ts) mutant of Escherichia coli K12

Abstract

In dnaK7(Ts) mutant cells, scission of DNA strands occurred after temperature shift up. When cells at 30 degrees C were labeled with [3H]-thymidine and then shifted to 46 degrees or 49 degrees C for 20 min, the profiles of sedimentation of their cellular DNA in an alkaline sucrose gradient revealed a decrease in the size of DNA to a quarter of that at 30 degrees C in the mutant, but not in wild-type cells. The level of manganese-containing superoxide dismutase (MnSOD) in the mutant was about twice that in wild-type cells, even at the permissive temperature, implying increased production of superoxide radical anion, which may cleave DNA strands directly or indirectly in the mutant. Moderate increase in the MnSOD level on temperature shift up was observed in both strains. These results indicated that some components of the DnaK protein participate in protection of cellular membrane functions from thermal damage resulting from elevated production of the superoxide anion radical.