Mutations simultaneously affecting endonuclease II and exonuclease III in Escherichia coli.
We studied mutants of E. coli originally identified as being deficient in either endonuclease II (deoxyribonucleate oligonucleotidohydrolase, EC 3.1.4.30) or exonuclease III [deoxyribonucleate (double-stranded) 5'-nucleotidohydrolase, EC 3.1.4.27] activity. Twelve independently derived mutants were tested, including three new endonuclease II mutants. Deficiency of one enzyme was always accompanied by deficiency of the other. Furthermore, temperature-sensitivity of one activity was always accompanied by temperature-sensitivity of the other, and the enzymes were co-purified. The results suggested a physical association between exonuclease III and endonuclease II, which may be of advantage in the excision-repair of DNA. A thermolabile endonuclease II was purified from one of the new mutants, indicating that it had an altered structural gene. This mutation, and all similar ones mapped by genetic transduction, was located between the pncA and aroD genes on the E. coli chromosome. One mutant had a prolonged generation time, an increased sensitivity to the alkylating agents methyl-methanesulfonate and mitomycin C, and a decreased plating efficiency for bacteriophage lambda, but no marked sensitivity to ultraviolet or gamma-irradiation. Its enzymatic and biological abnormalities were simultaneously revertible, suggesting they were caused by a single mutation. These results suggested a role for these enzymes in normal cell growth processes and in the repair of alkylation damage.
D002874:Chromosome Mapping; D003090:Coliphages; D003851:Deoxyribonucleases / Q000096:biosynthesis*; D004720:Endonucleases / Q000096:biosynthesis*; D004926:Escherichia coli / Q000201:enzymology*; D005092:Exonucleases / Q000096:biosynthesis*; D005796:Genes; D008040:Genetic Linkage; D009154:Mutation; D010641:Phenotype; D013696:Temperature; D014161:Transduction, Genetic
D003851:Deoxyribonucleases; D004720:Endonucleases; D005092:Exonucleases