Properties of a human lymphoblast AP-endonuclease associated with activity for DNA damaged by ultraviolet light, gamma-rays, or osmium tetroxide.
An endonuclease activity for UV-irradiated DNA, gamma-irradiated DNA, and OsO4-treated DNA that was partially purified from human lymphoblasts was found to have associated with it an endonuclease activity for partially depurinated DNA. When this apurinic endonuclease (Endo A) was characterized and compared with the cells' major apurinic endonuclease (Endo B), several notable differences were observed. (1) Endo A bound to oxidized DNA-Sepharose under conditions where Endo B did not. (2) Endo A did not require Mg2+, retaining full activity in 10 mM ethylenediamine-tetraacetic acid, while Endo B showed an absolute requirement for Mg2+. (3) Whereas the nicks made in depurinated DNA by Endo B were efficient priming sites for Escherichia coli polymerase I, those made by Endo A were not. Further characterization of the nicks indicated that Endo A incises depurinated DNA 3' to apurinic sites, leaving 3'-terminal deoxyribose, a poor priming site for DNA synthesis. Endo A action on UV-irradiated DNA produced nicks that resembled those it made in depurinated DNA, suggesting that the UV endonuclease activity acts through an apurinic/apyrimidinic site intermediate.
D002460:Cell Line; D004247:DNA / Q000528:radiation effects*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D043223:Deoxyribonuclease IV (Phage T4-Induced); D004706:Endodeoxyribonucleases / Q000302:isolation & purification / Q000378:metabolism*; D029968:Escherichia coli Proteins; D005720:Gamma Rays; D006801:Humans; D007700:Kinetics; D007945:Leukemia, Lymphoid / Q000201:enzymology*; D009992:Osmium / Q000633:toxicity*; D009993:Osmium Tetroxide / Q000633:toxicity*; D014466:Ultraviolet Rays
D029968:Escherichia coli Proteins; D009992:Osmium; D004247:DNA; D004706:Endodeoxyribonucleases; D043223:Deoxyribonuclease IV (Phage T4-Induced); C468972:endonuclease IV, E coli; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D009993:Osmium Tetroxide