Evidence for an imino intermediate in the DNA polymerase beta deoxyribose phosphate excision reaction

J Biol Chem. 1996 Jul 26;271(30):17811-5. doi: 10.1074/jbc.271.30.17811.


A recent study demonstrated that rat DNA polymerase beta (beta-pol) releases 5'-deoxyribose phosphate (dRP) termini from preincised apurinic/apyrimidinic DNA, a substrate generated during certain types of base excision repair. This catalytic activity resides within the amino-terminal, 8-kDa domain of beta-pol and occurs via beta-elimination as opposed to hydrolysis (Matsumoto, Y., and Kim, K. (1995) Science 269, 699-702). The latter finding suggested that the dRP excision reaction might proceed via an imine intermediate. In order to test this hypothesis, we attempted to trap beta-pol on preincised apurinic/apyrimidinic DNA using NaBH4 as the reducing agent. Both 8-kDa domain-DNA and intact beta-pol-DNA complexes were detected and identified by autoradiography coupled to immunoblotting. Our results indicate that the chemical mechanism of the beta-pol dRpase reaction does proceed through an imine enzyme-DNA intermediate and that the active site residue responsible for dRP release must therefore contain a primary amine.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • DNA Polymerase I / metabolism*
  • DNA Repair*
  • Imines / metabolism*
  • Molecular Sequence Data
  • Rats
  • Ribosemonophosphates / metabolism*


  • Imines
  • Ribosemonophosphates
  • 2-deoxyribose 5-phosphate
  • DNA Polymerase I