Viability of rep recA mutants depends on their capacity to cope with spontaneous oxidative damage and on the DnaK chaperone protein.
Replication arrests due to the lack or the inhibition of replicative helicases are processed by recombination proteins. Consequently, cells deficient in the Rep helicase, in which replication pauses are frequent, require the RecBCD recombination complex for growth. rep recA mutants are viable and display no growth defect at 37 or 42 degrees C. The putative role of chaperone proteins in rep and rep recA mutants was investigated by testing the effects of dnaK mutations. dnaK756 and dnaK306 mutations, which allow growth of otherwise wild-type Escherichia coli cells at 40 degrees C, are lethal in rep recA mutants at this temperature. Furthermore, they affect the growth of rep mutants, and to a lesser extent, that of recA mutants. We conclude that both rep and recA mutants require DnaK for optimal growth, leading to low viability of the triple (rep recA dnaK) mutant. rep recA mutant cells form colonies at low efficiency when grown to exponential phase at 30 degrees C. Although the plating defect is not observed at a high temperature, it is not suppressed by overexpression of heat shock proteins at 30 degrees C. The plating defect of rep recA mutant cells is suppressed by the presence of catalase in the plates. The cryosensitivity of rep recA mutants therefore results from an increased sensitivity to oxidative damage upon propagation at low temperatures.
Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique, Domaine de Vilvert, F-78352 Jouy en Josas Cedex, France.
M F
MF
Bredèche
S D
SD
Ehrlich
B
B
Michel
D004265:DNA Helicases / Q000502:physiology*; D004261:DNA Replication; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000254:growth & development; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins / Q000502:physiology*; D006861:Hydrogen Peroxide / Q000633:toxicity; D009154:Mutation; D010084:Oxidation-Reduction; D011938:Rec A Recombinases / Q000235:genetics*; D015534:Trans-Activators / Q000502:physiology*
D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D006861:Hydrogen Peroxide; D011938:Rec A Recombinases; C063842:dnaK protein, E coli; D004265:DNA Helicases