A ts mutant isolated from CHL cells: inhibition of DNA replication at nonpermissive temperature

A mutant Chinese hamster cell clone has been isolated from an S phase culture. This clone, ts154, could not replicate DNA at nonpermissive temperature (39 degrees C), although it synthesized DNA normally at permissive temperature (33 degrees C). After transfer to 39 degrees, ts154 cells replicated DNA semiconservatively through precisely two complete rounds and divided twice before they ceased DNA synthesis and further growth. The mutation in ts154 affects DNA replication specifically, as its rates of cellular transcription, its ability to support growth of vesicular stomatitis virus, and its ribo- and deoxyribonucleoside triphosphate pools were normal at 39 degrees. Cultures of ts154 resumed DNA replication within 6h when shifted from 39 degrees to 33 degrees, even after 78 h at the nonpermissive temperature. Upon return to 33 degrees, mutant cells achieved a normal steady-state rate of DNA synthesis by 10 h, suggesting that they had come to rest within G1 phase at 39 degrees. Resumption of DNA synthesis and growth after a shift-down to 33 degrees were sensitive to cycloheximide in ts154, and thus likely required synthesis of new protein.