Studies on DNA repair in Bacillus subtilis. II. Partial purification and mode of action of an enzyme enhancing the priming activity of γ-irradiated DNA

Abstract

A cellular factor which makes T7 DNA irradiated with γ-rays a better primer for Micrococcus DNA polymerase was partially purified by DEAE and phosphocellulose column chromatography and named “primer activating enzyme”. Sucrose density gradient sedimentation analysis was carried out to examine actions of one major active fraction that appeared by phosphocellulose chromatography. It was shown that this factor introduced new nicks in T7 DNA in addition to those introduced directly by γ-ray irradiation. This enzyme fraction also had an endonucleolytic activity towards DNA containing apurinic sites induced by heat treatment and had capacity to enhance the priming activity of heat- or methyl methansulfonate-treated DNA but affected very little that of ultraviolet-irradiated DNA. This enzyme had no effect on T7 DNA when it was not treated with the DNA-damaging agents. From these results we concluded that this enzyme may be analogous to the endonuclease II or apurinic site-specific endonuclease of Escherichia coli.