Repair of neocarzinostatin-induced deoxyribonucleic acid damage in human lymphoblastoid cells: possible involvement of apurinic/apyrimidinic sites as intermediates

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Abstract
Neocarzinostatin (NCS) [an antitumor agent] induces repair in a xeroderma pigmentosum lymphoblastoid line (XPA-3 cells), which is deficient in the ability to repair DNA damage induced with (acetoxyacetylamino)fluorence. Repair was demonstrated by the induction of repair synthesis and by the disappearance of NCS-induced single-strand breaks, and/or alkaline-labile sites in DNA. Estimation of NCS-induced repair patch size (based on the density shift induced in DNA by extensive shear after incubation of treated cells in medium with bromodeoxyuridine, or by calculation from the extent of restoration of DNA sedimentation profiles in alkaline sucrose gradients and the amount of repair synthesis measured by the BND cellulose method) indicated that only a few nucleotides were inserted per repaired region. NCS-treated bacteriophage T7 DNA requires incubation with alkaline phosphatase to make it a substrate for DNA polymerase I. NCS-reacted T7 DNA, even after phosphatase treatment, is not a substrate for a DNA polymerase .alpha. obtained from human lymphoma cells. NCS-treated T7 DNA did serve as a substrate for the DNA polymerase .alpha. when incubated with an apurinic/apyrimidinic (AP) endonuclease, with associated 5''-3''-exonuclease activity. The results suggest that NCS-induced AP sites could be intermediates for the in vivo repair synthesis.