Evidence suggestive of compartmentalization of deoxyribonucleic acid-synthesizing systems in freeze-treated Bacillus subtilis

Freezing of Bacillus subtilis in liquid nitrogen results, upon thawing of the cells, in an enhanced deoxyribonucleoside triphosphate and reduced thymidine (Tdr) incorporation into cellular deoxyribonucleic acid (DNA). The DNA synthesized from thymidine triphosphate (TTP) was made by a “repair”-type system as determined by density transfer experiments. The mono- and diphosphate precursors were also incorporated by a “repair”-type synthesis. When Tdr was used as the radioactive precursor in the assay mixture, the product was only that expected from a semiconservative synthesis. Superlethal ultraviolet light exposure of the freeze-treated cells stimulated incorporation of phosphorylated precursors into DNA. Tdr uptake was greatly reduced by ultraviolet exposure, and only repair synthesis was observed. TTP and Tdr do not compete with one another in this system. The possibility that two DNA synthesizing systems exist in separate, non-mixing cellular compartments is considered.