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J Biol Chem, Vol. 274, Issue 20, 13741-13743, May 14, 1999

COMMUNICATION
Role of DNA Polymerase  in the Excision Step of Long Patch Mammalian Base Excision Repair

Grigory L. Dianov, Rajendra Prasad^¶, Samuel H. Wilson^¶, and Vilhelm A. Bohr

From the  NIA, National Institutes of Health, Baltimore, Maryland 21224 and the ^¶ NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709

The two base excision repair (BER) subpathways in mammalian cells are characterized by the number of nucleotides synthesized into the excision patch. They are the "single-nucleotide" BER pathway and the "long patch" (several nucleotides incorporated) BER pathway. Both of these subpathways involve excision of a damaged base and/or nearby nucleotides and DNA synthesis to fill the excision gap. Whereas DNA polymerase  (pol ) is known to participate in the single-nucleotide BER pathway, the identity of polymerases involved in long patch BER has remained unclear. By analyzing products of long patch excision generated during BER of a uracil-containing DNA substrate in mammalian cell extracts we find that long patch excision depends on pol . We show that the excision of the characteristic 5'-deoxyribose phosphate containing oligonucleotide (dRP-oligo) is deficient in extracts from pol  null cells and is rescued by addition of purified pol . Also, pol -neutralizing antibody inhibits release of the dRP-oligo in wild-type cell extracts, and the addition of pol  after inhibition with antibody completely restores the excision reaction. The results indicate that pol  plays an essential role in long patch BER by conducting strand displacement synthesis and controlling the size of the excised flap.

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