title: An enzyme activity from Escherichia coli that attacks single-stranded deoxyribopolymers and single-stranded deoxyribonucleic acid containing apyrimidinic sites.
abstract: We have isolated an enzyme activity from extracts of Escherichia coli that catalyzes the hydrolysis of phosphodiester bonds in the single-stranded deoxyribopolymer (dU.[3H]dT)(2000) containing depyrimidinated sites created by enzymatic removal of uracil with uracil-DNA glycosylase. Nondepyrimidinated polymer is not degraded by the enzyme, nor is the depyrimidinated polymer degraded after reduction of apyrimidinic sites with sodium borohydride. The enzyme also degrades circular M13 DNA containing uracil or denatured phage PBS2 DNA (which naturally contains uracil) only after the removal of uracil from these substrates by uracil-DNA glycosylase. Undamaged or depurinated duplex PM2 or ColE1 DNA and undamaged M13 single-stranded DNA are not attacked by the enzyme. The activity sediments in glycerol gradients with a relative s value of 4.2 S and has a Stokes radius of 31 A, yielding a calculated Mr approximately 56,000. The fractional ratio (f/f0) is calculated at 1.25. The enzyme has no requirement for any known cofactors and is insensitive to inhibition by p-(chloromercuri)benzoate. Activity is inhibited in the presence of adenosine 5'0-triphosphate (ATP), tRNA or high ionic strength and is slightly stimulated by MnCl2 or CaCl2.
mesh_terms: D004260:DNA Repair; D004277:DNA, Single-Stranded; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D043223:Deoxyribonuclease IV (Phage T4-Induced); D003851:Deoxyribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D004720:Endonucleases / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D013379:Substrate Specificity
chemical_list: D004277:DNA, Single-Stranded; D029968:Escherichia coli Proteins; D003851:Deoxyribonucleases; D004720:Endonucleases; D043223:Deoxyribonuclease IV (Phage T4-Induced); C468972:endonuclease IV, E coli; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase