Identification and initial characterisation of a pyrimidine dimer UV endonuclease (UV endonuclease beta) from Deinococcus radiodurans; a DNA-repair enzyme that requires manganese ions.
An endonuclease that incises lightly ultraviolet-irradiated supercoiled plasmid DNA was identified in cell-free extracts of Deinococcus radiodurans R1 wild-type. The endonuclease was absent from strains mutant in the uvsC, uvsD or uvsE genes identifying it as 'UV endonuclease beta' responsible for the initial incision step of one excision-repair pathway for the removal of pyrimidine dimers from D. radiodurans DNA in vivo. The enzyme was purified free from contaminating nuclease activities and was partially characterised. The enzyme has an apparent molecular weight of 36 000, is ATP-independent, caffeine-insensitive and is inactivated by N-ethylmaleimide. It also has a novel requirement for manganese ions distinguishing it from all other known DNA-repair enzymes.
D002850:Chromatography, Gel; D004260:DNA Repair / Q000528:radiation effects; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000528:radiation effects; D004278:DNA, Superhelical / Q000378:metabolism / Q000528:radiation effects; D043224:Deoxyribonuclease (Pyrimidine Dimer); D004587:Electrophoresis, Agar Gel; D004706:Endodeoxyribonucleases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000302:isolation & purification*; D005798:Genes, Bacterial; D008345:Manganese / Q000494:pharmacology*; D008837:Micrococcus / Q000201:enzymology / Q000235:genetics* / Q000528:radiation effects; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000302:isolation & purification*; D009154:Mutation; D009699:N-Glycosyl Hydrolases; D010957:Plasmids; D014466:Ultraviolet Rays
D004269:DNA, Bacterial; D004278:DNA, Superhelical; D009097:Multienzyme Complexes; D008345:Manganese; C032219:UV endonuclease; D004706:Endodeoxyribonucleases; D043224:Deoxyribonuclease (Pyrimidine Dimer); D009699:N-Glycosyl Hydrolases