Coexpression of two thermosensitive defects in a Chinese hamster cell line: Manifestation of a G1 block associated with a mitosis defect

Abstract

Asynchronous cultures of ts 12, an anchorage-dependent derivative of the thermosensitive Chinese hamster cell line ts111, show a rapid drop in [ 3H]thymidine incorporation with accumulation of the cells in the G1 and in the G2 phases of the cycle, when shifted from 34.5 to 39.4 °C. Shift-up experiments carried out after either isoleucine deprivation or synchronization at 39.4 °C, locate the execution point of a ts function in late G1 (2.5–3 h before S). However, stimulation of proliferation of a high density-arrested population allows a fraction of the cells to enter S. In addition to the G1 ts defect, ts12 expresses a slight cytokinesis defect at 39.4 °C (8–15% binucleate cells). The results suggest that altered processes are taking place at a post-metaphasic stage during the first hours after the shiftup. When populations are synchronized by a thymidine block and released at 39.4 °C, multinucleate cells in addition to binucleate cells are observed. Part of these multinucleate cells result from abnormal karyokinesis without inhibition of cytokinesis. Evidence is presented suggesting that excess thymidine allows the re-expression of the multinucleation phenotype of ts111.