Floros J, Ashihara T, Baserga R
Cell Biol Int Rep. 1978 May;2(3):259-69. doi: 10.1016/0309-1651(78)90006-1.
ts 13 cells are a temperature-sensitive (ts) mutant of BHK cells that are known to arrest in G1 when shifted to the nonpermissive temperature. We have determined the entry into S of ts13 cells in five different growth conditions, namely: 1) quiescent, sparse cultures stimulated to proliferate by serum. 2) Quiescent, dense cultures stimulated by serum. 3) Quiescent, sparse cultures stimulated by trypsinization and replating. 4) Quiescent, dense cultures stimulated by trypsinization and replating. 5) Mitotic cells collected by mitotic detachment. For each different growth condition we have also determined the execution point of the mutant function, i.e. the time at which a shift-up to the nonpermissive temperature no longer prevents the entry of cells into S. The median time of entry into S and the execution point varied in different growth conditions, but the distance between the median execution point and the median time of entry into S was remarkably constant, i.e. 3.2 hr. In addition we have fused ts 13 cells cells with chick erythrocytes and studied the ability of ts13 cells in heterokaryon formation to induce DNA synthesis in chick nuclei. Although ts13 cells can induce DNA synthesis in chick nuclei at the permissive temperature, they fail to do so when fused and stimulated at the nonpermissive temperature of 39.5 degrees C.
ts13细胞是BHK细胞的温度敏感(ts)突变体,已知当转移到非允许温度时会停滞在G1期。我们已经确定了ts13细胞在五种不同生长条件下进入S期的情况,即:1)静止、稀疏的培养物,通过血清刺激增殖。2)静止、密集的培养物,通过血清刺激。3)静止、稀疏的培养物,通过胰蛋白酶消化和重新铺板刺激。4)静止、密集的培养物,通过胰蛋白酶消化和重新铺板刺激。5)通过有丝分裂脱离收集的有丝分裂细胞。对于每种不同的生长条件,我们还确定了突变功能的执行点,即转移到非允许温度后不再阻止细胞进入S期的时间。进入S期的中位时间和执行点在不同生长条件下有所不同,但中位执行点与进入S期的中位时间之间的距离非常恒定,即3.2小时。此外,我们将ts13细胞与鸡红细胞融合,并研究了ts13细胞在异核体形成中诱导鸡细胞核DNA合成的能力。尽管ts13细胞在允许温度下可以诱导鸡细胞核DNA合成,但当在39.5℃的非允许温度下融合并刺激时,它们无法做到这一点。