Mutants temperature sensitive for proliferation or survival were isolated from an untransformed diploid clone of fibroblastic rat cells (3Y1), according to an isolation protocol that selected for mutants defective at 38.5 degrees C (selection temperature) in undergoing the transition from quiescent to proliferating state while maintaining viability at 38.5 degrees C. Of the 108 temperature-sensitive clones isolated, 32 were examined for survival in sparse cultures at 39.8 degrees C (nonpermissive temperature) and classified into four classes. Results of temperature shift-up experiments suggest that functions defective in 11 of the 32 mutants are necessary not only for changing from the quiescent to proliferating state but also for maintenance of the proliferating state. Of the 32 mutants, 17 were assigned to eight complementation groups. Results of the physiological characterization of the representative mutants of each of the eight complementation groups are presented.