Mutation and killing of Escherichia coli expressing a cloned Bacillus subtilis gene whose product alters DNA conformation.
Expression of the Bacillus subtilis gene coding for SspC, a small, acid-soluble protein, caused both killing and mutation in a number of Escherichia coli B and K-12 strains. SspC was previously shown to bind E. coli DNA in vivo, and in vitro this protein binds DNA and converts it into an A-like conformation. Analysis of revertants of nonsense mutations showed that SspC caused single-base changes, and a greater proportion of these were at A-T base pairs. Mutation in the recA gene abolished the induction of mutations upon synthesis of SspC, but the killing was only slightly greater than in RecA+ cells. Mutations in the umuC and umuD genes eliminated most of the mutagenic effect of SspC but not the killing, while the lexA mutation increased mutagenesis but did not appreciably affect the killing. Since there was neither killing nor mutation of E. coli after synthesis of a mutant SspC which does not bind DNA, it appears likely that the binding of wild-type SspC to DNA, with the attendant conformational change, was responsible for the killing and mutation. A strain containing the B. subtilis gene that is constitutive for the RecA protein at 42 degrees C showed a lower frequency of mutation when that temperature was used to induce the RecA protein than when the temperature was 30 degrees C, where the RecA level is low, suggesting that at the elevated temperature the high RecA level could be inhibiting binding of the B. subtilis protein to DNA.
Setlow
J K
JK
Biology Department, Brookhaven National Laboratory, Upton, New York 11973.
Randesi
M
M
Adams
J G
JG
Setlow
B
B
Setlow
P
P
D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D003001:Cloning, Molecular; D004259:DNA-Directed DNA Polymerase; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000502:physiology*; D029968:Escherichia coli Proteins; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D016296:Mutagenesis / Q000187:drug effects*; D009690:Nucleic Acid Conformation / Q000187:drug effects; D011938:Rec A Recombinases / Q000235:genetics; D012697:Serine Endopeptidases; D012808:Sigma Factor; D014157:Transcription Factors; D000099160:LexA Repressor Protein
D001426:Bacterial Proteins; D004259:DNA-Directed DNA Polymerase; D029968:Escherichia coli Proteins; D007544:Isopropyl Thiogalactoside; D011938:Rec A Recombinases; D012697:Serine Endopeptidases; D012808:Sigma Factor; D014157:Transcription Factors; D000099160:LexA Repressor Protein; C092468:spoIIR protein, Bacillus subtilis; C030069:spore-specific proteins, Bacillus; C055218:UmuD protein, E coli