Hep-2 cells were synchronized by a double thymidine block and infected with poliovirus type I (Mahoney) in hourly intervals after release from the second thymidine block. The S-phase is not prevented by a poliovirus infection but, with cells infected 0-4 hours after release, an increase of its duration is found. With an infection 5 hours and later after release, the duration of the S-phase is not different from that of an uninfected, synchronized control culture. DNA synthesis itself is slower early in S-phase and gets inhibited up to 75 per cent late in S-phase. All cultures show the first signs of a CPE 3.5 hours p.i. and, in spite of CPE, the cells continue to synthesize DNA. In infected cells a slightly higher peak of mitotic cells compared to control cultures is found. The time point of the mitotic peak is dependant of the time of infection and seems no longer controlled by the cell cycle. The mitotic indices are similar for all cultures infected at different times after release. When the cells are infected early after release CPE appears before mitosis and prevents the cells from entering mitosis. Cells which are infected towards the end of the S-phase finish mitosis normally before they exhibit characteristics of CPE. Extent and kinetics of poliovirus RNA synthesis and yield of virus progeny are not altered by the cell cycle.