Isolation of protease-proficient, recombinase-deficient recA mutants of Escherichia coli K-12.
We isolated recA mutants with altered protease activity and then examined recombinase activity to determine whether the protease and recombinase functions of the RecA protein of Escherichia coli are separable. We found five mutants that had moderately strong constitutive RecA protease activity but no recombinase activity above the delta recA strain background, the first clear-cut examples of mutants of this class, designated Prtc Rec-. We also isolated 65 mutants that were protease-defective toward the LexA repressor and found that all of them were also recombinase deficient. Four of these mutants retained both partial recombinase activity and partial inducible protease activity. The recombinase-defective mutants were much more sensitive than the recA+ strain to crystal violet, kanamycin, and chloramphenicol, indicating altered membrane permeability. The recA (Prtc Rec-) mutants had a subtle alteration in protease specificity, all being defective in spontaneous induction of phages lambda imm434 and 21. They differed from Prtc Rec+ mutants of comparable or even weaker constitutive protease strength, all of which showed dramatic spontaneous induction of these prophages. However, treating a Prtc Rec- mutant with mitomycin C resulted in significant prophage induction. Thus, the RecA proteins of the Prtc Rec- mutants have constitutive protease activity toward the LexA repressor, but have only DNA damage-activable protease activity toward phage repressors. UV-induced mutagenesis from his to his+ was studied for one Prtc Rec- mutant, and induced mutation frequencies as high as those for the recA+ strain were found despite the absence of recombinase activity.
Tessman
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Peterson
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D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D010582:Bacteriophage lambda / Q000235:genetics; D002463:Cell Membrane Permeability; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification / Q000528:radiation effects; D007700:Kinetics; D008242:Lysogeny; D016685:Mitomycin; D008937:Mitomycins / Q000494:pharmacology; D009154:Mutation; D010447:Peptide Hydrolases / Q000235:genetics*; D011938:Rec A Recombinases / Q000235:genetics*; D011995:Recombination, Genetic; D014466:Ultraviolet Rays
D000903:Antibiotics, Antineoplastic; D008937:Mitomycins; D016685:Mitomycin; D004254:DNA Nucleotidyltransferases; D011938:Rec A Recombinases; D010447:Peptide Hydrolases