Base Replacement Mechanisms for the Repair of Unnatural and Damaged Bases in DNA

  • Zvi Livneh
  • Joseph Sperling
Part of the NATO Advanced Study Institutes Series book series (NSSA, volume 40)


A purine insertase activity was found in extracts from E. coli H502. This activity brings about the insertion of adenine and of guanine into the appropriate apurinic sites in DNA, using dATP and dGTP as the purine donors, in a Mg++-dependnet reaction. Using extracts prepared from E. coli strains KMBL 1719 and 1720 we found a Mg++-dependent insertion of the free purine bases into depurinated DNA. It is possible that induced functions are involved in the process of insertion since active extracts could be prepared from E. coli strains (including AB3027 ap endo-poll-) only if they were grown in the presence of sublethal doses of alkylating agents.

A new method was developed for assaying base replacement mechanisms, based on the use of DNA with labeled AP sites. This method is currently used to look for a pyrimidine insertase activity in E. coli.


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  1. 1.
    T. Lindahl and B. Nyberg, Rate of depurination of native deoxyribonucleic acid, Biochemistry, 11: 3610 (1972).CrossRefGoogle Scholar
  2. 2.
    P.D. Lawley and P. Brookes, Further studies on the alkylation of nucleic acids and their constituent nucleotides, Biochem. J. 89: 127 (1963).CrossRefGoogle Scholar
  3. 3.
    T. Lindahl, DNA glycosylases, endonucleases for apurinic/ apyrimidinic sites and base excision repair, Prog. Nucl. Acid Res. Mol. Biol. 22: 135 (1979).CrossRefGoogle Scholar
  4. 4.
    W.G. Verly, F. Gossard and P. Crine, In vitro repair of apurinic sites in DNA, Proc. Natl. Acad. Sci. USA, 71: 2273 (1974).CrossRefGoogle Scholar
  5. 5.
    K. Bose, P. Karran and B. Strauss, Repair of depurinated DNA in vitro by enzymes purified from human lymphoblasts, Proc. Natl. Acad. Sci. USA, 75: 794 (1978).CrossRefGoogle Scholar
  6. 6.
    Z. Livneh, D. Elad and J. Sperling, Enzymatic insertion of purine bases into depurinated DNA in vitro, Proc. Natl. Acad. Sci. USA, 76: 1089 (1979).CrossRefGoogle Scholar
  7. 7.
    W.A. Deutsch and S. Linn, DNA binding activity from cultured human fibroblasts that is specific for partially depurinated DNA and that inserts purines into apurinic sites, Proc. Natl. Acad. Sci. USA, 76: 141 (1979).CrossRefGoogle Scholar
  8. 8.
    W.A. Deutsch and S. Linn, Further characterization of a depurinated DNA - purine base insertion activity from cultured human fibroblasts, J. Biol. Chem. 254: 12099 (1979).PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1981

Authors and Affiliations

  • Zvi Livneh
    • 1
  • Joseph Sperling
    • 1
  1. 1.Department of Organic ChemistryThe Weizmann Institute of ScienceRehovotIsrael

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