Chain elongation and joining of DNA synthesized during hydroxyurea treatment of Chinese hamster cells

Abstract

We have previously presented evidence that hydroxyurea treatment of synchronized G 1 Chinese hamster cells did not prevent the entry of cells into the DNA synthetic period but that the DNA synthesized during this period (in which total DNA synthesis was severely depressed) was quite small (Walters, R.A., Tobey, R.A. and Hildebrand, C.E. (1976) Biochem. Biophys. Res. Com. 69, 212–217). In view of the reported effects of hydroxyurea on deoxyribonucleoside metabolism and possible relationship to control of DNA replication (Bjursell, G. and Reichard, P. (1973) J. Biol. Chem. 248, 3904–3909 and Walters, R.A., Tobey, R.A. and Ratliff, R.L. (1973) Biochim. Biophys. Acta 319, 336–347), we examined the fate of DNA synthesized during and shortly after hydroxyurea treatment to determine if this DNA exhibited any kinetic behavior which might be an indicator of aberrant synthesis. We found that, upon hydroxyurea removal, DNA grew at a linear rate of 0.98 ± 0.12 · 10 6 dalton/min ( 0.98 ± 0.12 μ m/min ) for about 2.3 h. Beginning at 2.3 h, DNA with a molecular weight approx. 1.4 · 10 8 was very rapidly integrated into bulk DNA of ≥ 3.5 · 10 8 daltons. The apparent growth rate of the 1.4 · 10 8 dalton DNA was approx. 10.6 μm/min. The data suggest that, at least for this DNA, joining into bulk DNA required one-third to one-half of the S period to begin and, once begun, occurred very rapidly. The possibility of integration of replicon clusters is considered.