Comparison of apurinic DNA-binding protein from an ataxia telangiectasia and a HeLa cell line. Evidence for an altered processing of apurinic/apyrimidinic endonuclease.
The major apurinic (AP) DNA-binding protein was purified from a HeLa cell line and from the SV40-transformed cell line AT5BIVA derived from a patient with the repair deficiency syndrome ataxia telangiectasia (AT). This protein appears to be identical with the major cellular apurinic/apyrimidinic endonuclease. The two endonucleases differ in their molecular weight (HeLa, 37,600; AT, 38,900) and their dissociation equilibrium constant for AP sites (HeLa, 7.8 X 10(-11) M; AT, 28.3 X 10(-11) M). These variances might be the consequence of a different post-translational modification. Evidence for this interpretation stems from the observation that the AP DNA binding activity of AP endonuclease, as measured in a glass-fiber filter binding assay, is inactivated upon incubation with snake venom phosphodiesterase and that the AP endonuclease from AT cells in 5-10-fold more sensitive than the HeLa enzyme. For both enzymes, the diesterase treatment leads to the formation of a protein of Mr 35,500 which might be the unmodified precursor of AP endonuclease. The loss of AP DNA binding does not reduce but rather increases the catalytic activity of AP endonuclease when measured at excess substrate concentration.
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Kuhnlein
D001260:Ataxia Telangiectasia / Q000378:metabolism*; D002460:Cell Line; D002472:Cell Transformation, Viral; D002846:Chromatography, Affinity; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D004268:DNA-Binding Proteins / Q000032:analysis; D003720:Densitometry; D043223:Deoxyribonuclease IV (Phage T4-Induced); D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D006367:HeLa Cells / Q000032:analysis*; D006801:Humans; D008970:Molecular Weight; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D013539:Simian virus 40
D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D043223:Deoxyribonuclease IV (Phage T4-Induced); D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; C470790:phosphodiesterase I, snake venom; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase