Kinetics of thymidine incorporation into detergent-soluble DNA of mouse lymphocytes

We reported recently that splenocytes from concanavalin A-stimulated mice rapidly incorporated [3H]thymidine into non-mitochondrial DNA that was detergent soluble and distributed in size classes between 200 and 5000 base pairs. In this report we show that small [3H]thymidine-labeled oligonucleotides (less than 100 base pairs long) appear by 15 min. Subsequently, [3H]thymidine in small oligonucleotides diminishes as incorporation into larger size classes of detergent-soluble DNA occurs in a pattern that is stable for at least 3 hr. Although incorporation of [3H]thymidine into the oligonucleotides is not sensitive to aphidicolin or hydroxyurea, the appearance of [3H]thymidine in larger species is blocked by both drugs. These results indicate that the enzymatic process involved in synthesis of oligonucleotides is somehow distinct from the process involved in synthesis of larger detergent-soluble size classes. Synthesis of the latter may be replication related.