Constitutive and UV-mediated activation of RecA protein: combined effects of recA441 and recF143 mutations and of addition of nucleosides and adenine.
The recF143 mutant of Escherichia coli is deficient in certain functions that also require the RecA protein: cell survival after DNA damage, some pathways of genetic recombination, and induction of SOS genes and temperate bacteriophage through cleavage of the LexA and phage repressors. To characterize the role of RecF in SOS induction and RecA activation, we determined the effects of the recF143 mutation on the rate of RecA-promoted cleavage of LexA, the repressor of the SOS genes. We show that RecA activation following UV irradiation is delayed by recF143 and that RecF is specifically involved in the SOS induction pathway that requires DNA replication. At 32 degrees C, the recA441 mutation partially suppresses the defect of recF mutants in inducing the SOS system in response to UV irradiation (A. Thomas and R. G. Lloyd, J. Gen. Microbiol. 129:681-686, 1983; M. R. Volkert, L. J. Margossian, and A. J. Clark, J. Bacteriol. 160:702-705, 1984); we find that this suppression occurs at the earliest detectable phase of LexA cleavage and does not require protein synthesis. Our results support the idea that following UV irradiation, RecF enhances the activation of RecA into a form that promotes LexA cleavage (A. Thomas and R. G. Lloyd, J. Gen. Microbiol. 129:681-686, 1983; M. V. V. S. Madiraju, A. Templin, and A. J. Clark, Proc. Natl. Acad. Sci. USA 85:6592-6596, 1988). In contrast to the constitutive activation phenotype of the recA441 mutant, the recA441-mediated suppression of recF is not affected by adenine and nucleosides. We also find that wild-type RecA protein is somewhat activated by adenine in the absence of DNA damage.
Sassanfar
M
M
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Roberts
J
J
D000225:Adenine / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism*; D001761:Bleomycin / Q000494:pharmacology; D004261:DNA Replication; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics* / Q000528:radiation effects; D016153:Genes, Suppressor; D006358:Hot Temperature; D009154:Mutation; D009268:Nalidixic Acid / Q000494:pharmacology; D011938:Rec A Recombinases / Q000235:genetics*; D013014:SOS Response, Genetics; D012697:Serine Endopeptidases; D013997:Time Factors; D014466:Ultraviolet Rays; D000099160:LexA Repressor Protein
D000225:Adenine; D001426:Bacterial Proteins; D001761:Bleomycin; D009268:Nalidixic Acid; D011938:Rec A Recombinases; D012697:Serine Endopeptidases; D000099160:LexA Repressor Protein